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Using modified amino ethers of boric acid, porous polyurethanes are synthesized, the porous morphology of which is due to the peculiarities of the hyperbranched macromolecular architecture of amino ethers of boric acid. A change in the structure of amino ethers of boric acid by the introduction of terminal bulky substituents with hydroxyl groups contributes to a change in the size and pore size distribution of polyurethanes based on amino ethers of boric acid. It is shown that one of the reasons for the heat-sensitive vapor permeability of porous polyurethanes is the hydrophilic nature of the pore surface. Film samples of porous polyurethanes have proven to be an effective substrate for the targeted cultivation of connective tissue at the cellular level.

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.

Long considered both physiologic and inevitable, skin aging is a degenerative phenomenon whereby both intrinsic and environmental factors conspire to produce an authentic disease. The consequences of this disorder are many and varied, ranging from atrophy and fragility to defective repair to deficient immunity and vulnerability to certain infections. The pathobiologic basis for skin aging remains poorly understood. At a cellular level, stem cell dysfunction and attrition appear to be key events, and both genetic and epigenetic factors are involved in a complex interplay that over time results in deterioration of our main protective interface with the external environment. Past and current understanding of the cellular and molecular intricacies of skin aging provide a foundation for future approaches designed to thwart the aging phenotype. Herein, the authors provide a review of current insights into skin aging, including the mechanisms of skin aging, the role of stem cells in skin aging and the implications of skin aging for the microbiome and for the development of cancer. Conquest of the oft overlooked disease of skin aging should have broad implications that transcend the integument and inform novel approaches to retarding aging and age-related dysfunction in those internal organs that youthful skin was designed to envelop and safeguard.

Background of study: The strength of a straight punch, uppercut and hook is needed by a professional boxer to knock down his opponent. Therefore, in the training program for a boxer, it is necessary to analyze the biomechanical characteristics and bio motoric components, which influence its, strength, endurance, and speed by not ignoring psychological factors and the degeneration process that occurs. Degeneration is a natural process, which occurs in every individual, from the cellular level to the level of movement. It functions since 30 years of age characterized by the disappearance of the ability of cells and tissues to repair and replace themselves and maintain normal structure, as well as resulting a decrease in all body functions for 1% every year Methodology: This article is a qualitative description with a literature study which analyzes various theories by experts in bio motoric components, degeneration processes, and psychological factors in the form of anxiety. Result: A balance between physical can slow the degeneration process, psychological, and environmental factors including the life style of a boxer, the factors of strength, endurance, speed, and psychological factors in the form of anxiety influence each other, both directly and indirectly against peak performance in the achievement of a boxer. Conclusion: Periodic measurements and evaluations of bio motoric components and mental training have to be considered, so that during the golden age, boxers can achieve optimally

Trabeculectomy is the most commonly performed surgery for the definitive treatment of glaucoma. Despite its high resolvability, the postoperative period requires high caution so that excessive filtration or scarring does not occur. This paper aimed to research alternative options to those most used as healing modulators, mitomycin C (MMC) and 5-fluorouracil, commonly associated with complications. This systematic review used the PubMed and SciELO databases, covering publications from 1972 to 2019. A total of 31 substances and methods were analyzed. Some, such as anti-VEGF, glucocorticoids and betatherapy, did not show results statistically superior to those of MMC. Others, such as the enzyme α5β1-integrin and Ologen®, demonstrated efficacy and safety at least similar to that of this drug. In conclusion, further research is still needed for drugs that lead to the same results as mitomycin, but with fewer side effects. More recent studies have focused on technologies that increase communication between target tissues and antifibrotic molecules at the cellular level, being a promising bet for the future.

Nitrogen (N) is the most abundant mineral elements in plants, and the application of inorganic N fertilizer makes huge contribution to the crop production and global food security. However, low N use efficiency (NUE) and overapplication of N fertilizers causes ever-growing environmental problems. Understanding the molecular mechanisms of N sensing and signalling in plants will provide molecular basis for NUE improvement of crops. Forward genetics screening and functional analysis have characterized the NRT1.1-NLP centered N signalling pathway at the cellular level. With the incorporation of systems biology approaches, a preliminary N regulatory network has been delineated. Meanwhile, long-distance N signalling has also been unveiled at the whole plant level. This review highlights most recent understanding of the N signalling network in plants, and also discusses how to further integrate hierarchical regulation of N signalling in plants.

Single-cell RNA sequencing (scRNA-seq), a method of transcriptome sequencing at the single-cell level, has recently emerged as a revolutionary technology in the field of biomedical research. Compared to conventional gene expression profiling in bulk, scRNA-seq resolves biological differences among individual cells and enables the identification of rare cell populations that are easily overlooked. This review introduces the method of scRNA-seq, summarizes its applications in the field of cardiovascular disease research, and discusses existing limitations and prospects for future applications.

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.

Summary There are a multitude of resistance strategies that microbes can apply to avoid inhibition by antimicrobials. One of these strategies is the enzymatic modification of the antibiotic, in a process generally termed inactivation. Furthermore, some microorganisms may not be limited to the mere inactivation of the antimicrobial compounds. They can continue by further enzymatic degradation of the compounds' carbon backbone, taking nutritional and energetic advantage of the former antibiotic. This driving force to harness an additional food source in a complex environment adds another level of complexity to the reasonably well‐understood process of antibiotic resistance proliferation on a single cell level: It brings bioprotection into play at the level of microbial community. Despite the possible implications of a resistant community in a host and a lurking antibiotic failure, knowledge of degradation pathways of antibiotics and their connections is scarce. Currently, it is limited to only a few families of antibiotics (e.g. β‐lactams and sulfonamides). In this article, we discuss the fluctuating nature of the relationship between antibiotic resistance and the biodegradation of antibiotics. This distinction mainly depends on the genetic background of the microbe, as general resistance genes can be recruited to function in a biodegradation pathway.

Perfluoroalkyl substances (PFASs) are persistent chemicals in the environment. So far, little is known about their uptake potential in wetland plants. Here, we investigated the uptake and translocation of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) in eight common wetland plants, namely

The ability to assess the responses of cells exposed to nanoparticles at the single cell level will lead to mechanistic understanding of their toxicity and safety. Here, we demonstrated the heterog...

Screening for endocrine disrupting properties at the molecular and cellular level is developing rapidly, but can epidemiology bridge the gap to human health impact? Reviews by the World Health Organization and the United States Endocrine Society listed diseases which may be related to endocrine disrupting chemicals (EDCs). Much of the evidence relating these diseases to EDCs is from animal and epidemiological studies, many with significant weaknesses. What human health data sources are available? Some examples are provided from Nordic countries, Denmark, France, Switzerland, and the United States. Health register shortcomings are noted, including diagnostic criteria and "signal-to-noise" ratio (high background incidence rates). Issues with exposure assessment (human biomonitoring), data governance (FAIR principles), legislative hurdles, and patient consent are also illustrated. For all the above reasons, it is clear why generating reliable long-term data for human diseases putatively associated with EDCs has not yet been achieved and thus why it is difficult to bridge the gap between molecular/cellular/animal toxicity data and human health risks of EDCs. This will require international cooperation, sustained support, and public acceptance.

Summary This paper summarizes the mitigation strategies for the thermal runaway of lithium-ion batteries. The mitigation strategies function at the material level, cell level, and system level. A time-sequence map with states and flows that describe the evolution of the physical and/or chemical processes has been proposed to interpret the mechanisms, both at the cell level and at the system level. At the cell level, the time-sequence map helps clarify the relationship between thermal runaway and fire. At the system level, the time-sequence map depicts the relationship between the expected thermal runaway propagation and the undesired fire pathway. Mitigation strategies are fulfilled by cutting off a specific transformation flow between the states in the time sequence map. The abuse conditions that may trigger thermal runaway are also summarized for the complete protection of lithium-ion batteries. This perspective provides directions for guaranteeing the safety of lithium-ion batteries for electrical energy storage applications in the future.

Neurotransmitters maintain physiological condition and behaviour of the brain and body. Studies on transmitting mechanisms and change in concentration of particular neurotransmitters provide knowledge about how these substances enhance our ability to interpret complex neural pathophysiology and thus advancing us towards developing new therapeutic and diagnostic intervention. Since the role of neurotransmitters in the cognitive processes are much complex, therefore their detection-based analysis leads to expand our understanding on their functions and early disease detection. One of the most crucial aspects of neuroscience is developing profound understanding of neurotransmitter release kinetics. As compared to other techniques, employing nanobiosensors for studying neurotransmission and dynamic changes in neurons exhibit relatively fast, accurate and significant results where other techniques generally fail. Using advanced techniques, high sensitivity and specificity also furnish precise information of neurotransmitter transport at the cellular level. Over the past few years, the advanced nanomaterials in combination of various biomolecules and distinct polymers are widely used in development of biosensing platforms for in-vivo and in-vitro point of care neurotransmitters detection. The review article is focused upon summarizing present status of nanobiosensors based neurotransmitter detection for clinical samples. This firstly describes the importance of neurotransmitters, conventional methods for neurotransmitters detection; need of developing advanced sensing methods and importance of nanomaterials to develop these sensing platforms. Secondly, current status of developed electrochemical and optical nanosensors for different neurotransmitters detection is elaborated in detail. The reported work extensively explains the importance of nano based sensing platforms for understating of neurotransmitters detection. Last section of the review summarized the future perspectives of nanosensors in clinical set-up for neurotransmitter detection.

New phenylethanoid, glucooleoacteoside (2), along with 18 known compounds were isolated from the aqueous extract of Osmanthus fragrans. The structure of new compound was elucidated on the base of IR, HR-ESI-MS, 1D NMR, 2D NMR and DEPT analysis. Bioactivity investigation into human dermal fibroblasts (HDF) proliferation, type I collagen expression, and MMP-1 expression were examined with six phenylethanoids. We found that compounds 1, 3, 4, 7, and 9 showed moderate bioactivities. In conclusion, we demonstrated that the aqueous extract of flower of O. fragrans contained phenylethanoid-rich constituents which revealed bioactivities at the cellular level of HDFs.

Recent advances in food packaging materials largely rely on nanotechnology structuring. Owing to several unique properties of nanostructures that are lacking in their bulk forms, the incorporation of nanostructures into packaging materials has greatly improved the performance and enriched the functionalities of these materials. This review focuses on the functions and applications of widely studied nanostructures for developing novel food packaging materials. Nanostructures that offer antimicrobial activity, enhance mechanical and barrier properties, and monitor food product freshness are discussed and compared. Furthermore, the safety and potential toxicity of nanostructures in food products are evaluated by summarizing the migration activity of nanostructures to different food systems and discussing the metabolism of nanostructures at the cellular level and in animal models.

The powerful analgesic effects of opioid drugs have captivated the interest of physicians and scientists for millennia, and the ability of opioid drugs to produce serious undesired effects has been recognized for a similar period of time (Kieffer and Evans, 2009). Many of these develop progressively with prolonged or repeated drug use and then persist, motivating particular interest in understanding how opioid drugs initiate adaptive or maladaptive modifications in neural function or regulation. Exciting advances have been made over the past several years in elucidating drug-induced changes at molecular, cellular, and physiologic scales of analysis. The present review will highlight some recent cellular studies that we believe bridge across scales and will focus on optical imaging approaches that put opioid drug action "under the microscope." SIGNIFICANCE STATEMENT: Opioid receptors are major pharmacological targets, but their signaling at the cellular level results from a complex interplay between pharmacology, regulation, subcellular localization, and membrane trafficking. This minireview discusses recent advances in understanding the cellular biology of opioid receptors, emphasizing particular topics discussed at the 50th anniversary of the International Narcotics Research Conference. Our goal is to highlight distinct signaling and regulatory properties emerging from the cellular biology of opioid receptors and discuss potential relevance to therapeutics.

Ultrasound molecular imaging has been developed in the past two decades with the goal of non-invasively imaging disease phenotypes on a cellular level not depicted on anatomic imaging. Such techniques already play a role in pre-clinical research for the assessment of disease mechanisms and drug effects, and are thought to in the future contribute to earlier diagnosis of disease, assessment of therapeutic effects and patient-tailored therapy in the clinical field. In this review, we first describe the chemical composition and structure as well as the in vivo behavior of the ultrasound contrast agents that have been developed for molecular imaging. We then discuss the strategies that are used for targeting of contrast agents to specific cellular targets and protocols used for imaging. Next we describe pre-clinical data on imaging of thrombosis, atherosclerosis and microvascular inflammation and in oncology, including the pathophysiological principles underlying the selection of targets in each area. Where applicable, we also discuss efforts that are currently underway for translation of this technique into the clinical arena.

This article investigates metallization-induced recombination losses associated with the presence of silicon (Si) in the 1) rear passivation layers and 2) aluminum (Al) paste in fully screen-printed p -type passivated emitter and rear cell (PERC) solar cells. For 1), the rear sides of PERCs passivated with aluminum oxide films stacked with silicon nitride (SiN y ) films with various refractive indexes (n), as defined by the composition of the SiN y films, were fabricated. Si-rich SiN y films strongly degraded the implied open-circuit voltages of symmetrically passivated samples, that include cells. Elemental mapping analysis confirmed further metallization-induced losses at the cell level because of the formation of an Al-Si alloy within the passivation stacks, with the density of the alloy dependent on the refractive index. For 2), the rear sides of PERCs were metallized with Si-free Al paste or Al pastes that contain Si. High-Si paste had both good and bad effects: it produced thicker aluminum back surface fields but left partially-formed Al-Si alloy within the passivation films.

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.

The function of cells in their native habitat often cannot be reliably predicted from genomic data or from physiology studies of isolates. Traditional experimental approaches to study the function of taxonomically and metabolically diverse microbiomes are limited by their destructive nature, low spatial resolution or low throughput. Recently developed technologies can offer new insights into cellular function in natural and human-made systems and how microorganisms interact with and shape the environments that they inhabit. In this Review, we provide an overview of these next-generation physiology approaches and discuss how the non-destructive analysis of cellular phenotypes, in combination with the separation of the target cells for downstream analyses, provide powerful new, complementary ways to study microbiome function. We anticipate that the widespread application of next-generation physiology approaches will transform the field of microbial ecology and dramatically improve our understanding of how microorganisms function in their native environment. In this Review, Hatzenpichler et al. introduce next-generation physiology, which is a suite of new techniques that enable investigation into the phenotypes of individual cells in a non-destructive manner. Next-generation physiology complements genomics and culturing and provides new insights into microbiome function.

Foodborne pathogens have become ongoing threats in the food industry, whereas their rapid detection and classification at an early stage are still challenging. To address early and rapid detection, hyperspectral microscope imaging (HMI) technology combined with convolutional neural networks (CNN) was proposed to classify foodborne bacterial species at the cellular level. HMI technology can simultaneously obtain both spatial and spectral information of different live bacterial cells, while two CNN frameworks, U-Net and one-dimensional CNN (1D-CNN), were employed to accelerate the data analysis process. U-Net was used for automating cellular regions of interest (ROI) segmentation, which generated accurate cell-ROI masks in a shorter timeframe than the conventional Otsu or Watershed methods. The 1D-CNN was employed for classifying the spectral profiles extracted from cell-ROI and resulted in a higher accuracy (90%) than k-nearest neighbor (81%) and support vector machine (81%). Overall, the CNN-assisted HMI technology showed potential for foodborne bacteria detection.

The circadian clock is a genetic circuit that allows organisms to anticipate daily events caused by the rotation of the Earth. The plant clock regulates physiology at multiple scales, from cell division to ecosystem-scale interactions. It is becoming clear that rather than being a single perfectly synchronised timer throughout the plant, the clock can be sensitive to different cues, run at different speeds, and drive distinct processes in different cell types and tissues. This flexibility may help the plant clock to regulate such a range of developmental and physiological processes. In this review, using examples from the literature, we describe how the clock regulates development at multiple scales and discuss how the clock might allow local flexibility in regulation whilst remaining coordinated across the plant.

Bell pepper or sweet pepper (Capsicum annuum L. var. grossum) is highly susceptible to high-temperature stress (HT). Hence, search for donor across C. annuum for HT tolerance was undertaken by following Temperature Induction Response (TIR) technique. The induction and challenging temperature requirement for TIR screening were standardized in 1 d-old Capsicum seedlings. Forty Capsicum genotypes were evaluated based on the recovery growth (RG) and per cent reduction in recovery growth (%RRG) of the seedlings. The genotypes Punjab Guchhedar and Ajeet 1 were found to have maximum cellular level tolerance (CLT) to HT with higher RG and lower %RRG compared to the non-induced seedlings and HDC 75 was found to have minimum CLT. In order to confirm the findings, another experiment was conducted under managed stress and control conditions. Absolute yield obtained from both the environments were used to calculate stress tolerance indices such as heat susceptibility index, tolerance index, stress tolerance index, mean productivity, geometric mean productivity and yield stability index. Based on these tolerance indices, Punjab Guchhedar and Ajeet 1 were found to be highly tolerant and HDC 75 as highly susceptible. Further, the combined result of TIR and tolerance indices also gave the same result confirming Punjab Guchhedar and Ajeet 1 can be used as a donor for the future breeding programme aimed at evolving high-temperature-tolerant bell pepper cultivars. The result also confirms the fitness of TIR technique to screen Capsicum genotypes for tolerance to HT based on variability in acquired thermotolerance.

Background Multiple studies have reported adipose tissue reduction after the application of the High‐Intensity Focused Electromagnetic (HIFEM) field technology, yet cellular level evidence of the mechanisms has remained scarce. Objectives This study aims to verify or refute previous single‐study histological evidence and further investigates the proposed mechanism of apoptotic induction. Methods The thigh of two Large White pigs was treated with HIFEM for 30 minutes. Fat punch biopsies were collected from the application area before, immediately after, and 8 hours post‐treatment. Control samples were taken from the abdomen immediately after and 8 hours post‐treatment. Samples were analyzed for pro‐apoptotic DNA markers (BAX, BCL‐2, TXNIP, MMP9, TNF‐α), the levels of free fatty acids (FFA), and the pH levels of the adipose tissue. Results The levels of FFA in the treated adipose tissue increased on average by 127.1% immediately post‐treatment and by 134.1% 8 hours post‐treatment, indicating a rapid breakdown of lipids. The average recorded adipose pH changed from 7.30 ± 0.12 at baseline to 6.60 ± 0.07 immediately post‐treatment (P = .001) and to 7.19 ± 0.12 8 hours post‐treatment. The levels of BAX, TXNIP, MMP9, and TNF‐α increased post‐treatment while BCL‐2 decreased. Control samples showed constant levels of pH and pro‐apoptotic markers. The FFAs in the control samples were increased by 41.6%‐51.4%. Conclusion The changes in the levels of the pro‐apoptotic markers conformed to the previously reported elevated fat apoptosis post‐HIFEM treatments. These effects were accompanied by an increase in FFA levels, and by reduced pH levels, due to the increased acidity in the adipose tissue. Further research is required to explore the potential of nonthermal induction of apoptosis.

Motile cilia are a striking example of functional cellular organelle, conserved across all the eukaryotic species. Motile cilia allow swimming of cells and small organisms and transport of liquids across epithelial tissues. Whilst the molecular structure is now very well understood, the dynamics of cilia is not well established either at the single cilium level nor at the level of collective beating. Indeed, a full understanding of this requires connecting together behaviour across various lengthscales, from the molecular to the organelle, then at cellular level and up to the tissue scale. Aside from the fundamental interest in this system, understanding beating is important to elucidate aspects of embryonic development and a variety of health conditions from fertility to genetic and infectious diseases of the airways., a review of 11 pages, Submitted to Biochemical Society Transactions

Hybridomas are a commonly used, or even the only option, for laboratory study and pilot production of monoclonal antibodies (mAbs), which are crucial for both targeted therapy and biomedical study. A long-term culture of hybridomas will inevitably induce a heterogenization of the whole hybridoma population, resulting in a continuous growth of non-producing hybridomas. To overcome the limits of existing methods of screening heterogeneous hybridomas, in which the whole multi-round screening process is performed in multi-well plates or other discrete modules, this study presents a novel method in which all processing steps of a multi-round hybridoma screening are finished in a single microfluidic chip. This microfluidic chip comprehensively performs hybridoma trapping/proliferating/transferring and fluorescent identification of protein-antibody binding at single cell level. By performing a two-round screening of anti-CD45 mAb secreting hybridomas, the novel microfluidic chip was proved capable of screening several single high-producing hybridomas with minimum cell loss/human labor/time cost, and more importantly, enhanced accuracy and definite monoclonality, which is one of the most important properties of mAb production.

This tutorial review article is highlighting the fundamentals, instrumentation, and most recent trends of single-cell analysis by use of inductively coupled plasma-mass spectrometry (ICP-MS). It is shown that metals and hetero-elements being intrinsically present in cells, taken up by cells (for instance engineered metallic nanoparticles) or binding to a cell can be detected qualitatively by existing ICP-MS technologies on a single cell level. Adding a quantitative dimension to single-cell analysis by (laser ablation-) ICP-MS requires dedicated calibration and validation strategies, which are currently being established and are being critically discussed. In a tutorial part, the ICP-MS instruments, the measurement conditions, and the sample introduction and preparation techniques are introduced. The application section focuses on the state-of-the-art of single-cell analysis in suspension, using laser ablation or (imaging) mass cytometry. Finally, future trends are critically assessed.

The recent worldwide spreading of pneumonia-causing virus, such as Coronavirus, COVID-19, and H1N1, has been endangering the life of human beings all around the world. In order to really understand the biological process within a cell level and provide useful clues to develop antiviral drugs, information of Gram negative bacterial protein subcellular localization is vitally important. In view of this, a CNN based protein subcellular localization predictor called “pLoc_Deep-mGnet” was developed. The predictor is particularly useful in dealing with the multi-sites systems in which some proteins may simultaneously occur in two or more different organelles that are the current focus of pharmaceutical industry. The global absolute true rate achieved by the new predictor is over 98% and its local accuracy is around 94% - 100%. Both are transcending other existing state-of-the-art predictors significantly. To maximize the convenience for most experimental scientists, a user-friendly web-server for the new predictor has been established at http://www.jci-bioinfo.cn/pLoc_Deep-mGneg/, which will become a very useful tool for fighting pandemic coronavirus and save the mankind of this planet.

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.

The unexpected finding of an animal pigment, bilirubin, in the arils of the plant Strelitzia nicolai, has resulted in interest regarding bilirubin's formation, its ultimate function as well as its ultrastructure both in the humans and in the plant. The main objective of this study was to identify the structural components of seed arils of S. nicolai at a cellular level. Furthermore, this study compared the five different stages of aril maturation and tracked bilirubin formation within the cells. This was accomplished by stereomicroscopic, light microscopic and electron microscopical analysis. These included arils stained with toluidine blue and the modified Hall's staining to detect the presence of bilirubin. The results from the light, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed the different stages of aril development containing a large number of cellular contents. However, as the aril matures, the contents of the cell became denser. Additionally, arils became a more structured and rigid as they matured. The most noticeable feature of the arils was that of the colour change as they mature. It was clarified that the colour change of the arils was attributed to the production and accumulation of bilirubin within the cells.

Visually identifying pathogens favors rapid diagnosis at the point-of-care testing level. Here, we developed a microenvironment-sensitive aggregation-induced emission luminogen (AIEgen), namely IQ–Cm, for achieving fast discrimination of Gram-negative bacteria, Gram-positive bacteria and fungi by the naked-eye. With a twisted donor–acceptor and multi-rotor structure, IQ–Cm shows twisted intramolecular charge transfer (TICT) and AIE properties with sensitive fluorescence color response to the microenvironment of pathogens. Driven by the intrinsic structural differences of pathogens, IQ–Cm with a cationic isoquinolinium moiety and a membrane-active coumarin unit as the targeting and interacting groups selectively locates in different sites of three pathogens and gives three naked-eye discernible emission colors. Gram-negative bacteria are weak pink, Gram-positive bacteria are orange-red and fungi are bright yellow. Therefore, based on their distinctive fluorescence response, IQ–Cm can directly discriminate the three pathogens at the cell level under a fluorescence microscope. Furthermore, we demonstrated the feasibility of IQ–Cm as a visual probe for fast diagnosis of urinary tract infections, timely monitoring of hospital-acquired infection processes and fast detection of molds in the food field. This simple visualization strategy based on one single AIEgen provides a promising platform for rapid pathogen detection and point-of-care diagnosis., A simple AIEgen with three emission colors achieves rapid identification of Gram-negative bacteria, Gram-positive bacteria and fungi.

Constrained predictive control has emerged as a viable candidate for generating optimal real-time charging strategies for lithium ion batteries. Able to conform to hard constraints on problem variables, model predictive control (MPC) formulates an optimal 2-norm solution at each time step and can thus assure safe and reliable fast-charge operation. Standard MPC implementations make certain simplifying assumptions regarding future control actions beyond a specified control horizon. This paper demonstrates the potential gains that can be realized for the battery charge problem by relaxing these assumptions.

The existence of a magnetic compass system allowing animals to maintain the direction of their migratory journeys was first shown in birds about fifty years ago. Over the past half century, a large amount of indirect, mainly behavioral, data on the features of the avian magnetic compass and its relationship with the perception of light has been obtained. Based on these data, the dominant idea suggests that the primary magnetoreceptor in birds is located in the retina, and the most popular hypothesis concerning operation of such a magnetoreceptor is the radical pair model. According to this model, the primary receptor molecules that perceive the magnetic field are cryptochromes expressed in the neurons of the avian retina. It is assumed that cryptochrome molecules localized in photoreceptor cells may have an ordered orientation relative to the retina plane. When photons are absorbed in the short wavelength range of the spectrum, cryptochromes would produce a chemical response depending on the direction of the external magnetic field. The sensitivity of photochemical reactions involving radical pairs in cryptochromes to a magnetic field is confirmed both by theoretical calculations and experimentally. However, up to date there is no experimental data obtained on the molecular mechanisms of further transduction of signal induced by a magnetic field. The existing hypotheses on the possible nature of these mechanisms have some issues concerning their compatibility with the current concepts of the photoreception. These hypotheses ascribe special properties to the participants of the visual process both at the molecular and at the cellular level, which has not yet been proved experimentally. Thus, nowadays the science of magnetoreception is posed to consistently test the existing hypotheses on the molecular basis of avian magnetic compass.

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.

The recent worldwide spreading of pneumonia-causing virus, such as Coronavirus, COVID-19, and H1N1, has been endangering the life of human beings all around the world. In order to really understand the biological process within a cell level and provide useful clues to develop antiviral drugs, information of Gram positive bacteria protein subcellular localization is vitally important. In view of this, a CNN based protein subcellular localization predictor called “pLoc_Deep-mGpos” was developed. The predictor is particularly useful in dealing with the multi-sites systems in which some proteins may simultaneously occur in two or more different organelles that are the current focus of pharmaceutical industry. The global absolute true rate achieved by the new predictor is over 99% and its local accuracy is around 92% - 99%. Both are transcending other existing state-of-the-art predictors significantly. To maximize the convenience for most experimental scientists, a user-friendly web-server for the new predictor has been established at http://www.jci-bioinfo.cn/pLoc_Deep-mGpos/, which will become a very powerful tool for developing effective drugs to fight pandemic coronavirus and save the mankind of this planet.

The recent worldwide spreading of pneumonia-causing virus, such as Coronavirus, COVID-19, and H1N1, has been endangering the life of human beings all around the world. In order to really understand the biological process within a cell level and provide useful clues to develop antiviral drugs, information of virus protein subcellular localization is vitally important. In view of this, a CNN based virus protein subcellular localization predictor called “pLoc_Deep-mVirus” was developed. The predictor is particularly useful in dealing with the multi-sites systems in which some proteins may simultaneously occur in two or more different organelles that are the current focus of pharmaceutical industry. The global absolute true rate achieved by the new predictor is over 97% and its local accuracy is over 98%. Both are transcending other existing state-of-the-art predictors significantly. It has not escaped our notice that the deep-learning treatment can be used to deal with many other biological systems as well. To maximize the convenience for most experimental scientists, a user-friendly web-server for the new predictor has been established at http://www.jci-bioinfo.cn/pLoc_Deep-mVirus/.

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.

The lung consists of a plethora of cellular components working in concert to protect us against a variety of pathogens. This unique complexity poses a challenge to traditional transcriptomic approaches as averaging out signals across an intricate organ dampens individual cell type contributions. Technologies that allow transcriptomic level measurements at a single cell level, such as single cell RNA sequencing (scRNAseq) have enabled an unbiased survey of the organ during homeostasis and disease, while preserving intricate cellular heterogeneity. The utility of scRNAseq in respiratory science has been widely embraced over the past few years with key discoveries including the identification of rare cell types, new cell states associated with lung diseases, and a system wide expression of key severe acute respiratory syndrome coronavirus 2 (SARS-Cov2) receptors. Global single cell consortiums such as the Human Cell Atlas and LungMap have provided collaborative platforms that are crucial for large scale analysis, and they have emerged as important facilitators to further our understanding of lung biology. The rapid evolution of this technology will enable us to broaden the restraints of our knowledge leading to exciting discoveries that we currently see impossible.

The ability to deposit single cells in nonrestrictive two-dimensional and three-dimensional environments with high accuracy and reproducibility has proven invaluable for innovative cell and tissue research, and is now entering a new stage of maturity through systems engineering and next-generation designs. Coupling computer-aided design and computer-aided machining with laser direct write systems enables researchers to create reproducible biological models with micron-level precision. In particular, matrix-assisted pulsed-laser evaporation direct write system, and variations, has demonstrated this enhanced bottom-up approach to fabricating single-cell level constructs. This technology showcases the advantages of laser-assisted, nozzle-free, and contact-free printing that avoid probabilistic printing methods. In this chapter, we illustrate the importance of single-cell deposition and introduce laser-assisted bioprinting technology capable of micron-resolution and accuracy. This chapter will also review the post-deposition cell viability and mechanistics of laser-assisted transfer of biological materials.

SummaryReconstruction of shapes and sizes of three-dimensional (3D) objects from two-dimensional (2D) information is an intensely studied subject in computer vision. We here consider the level of single cells and nuclei and present a neural network-based SHApe PRediction autoencoder. For proof-of-concept, SHAPR reconstructs 3D shapes of red blood cells from single view 2D confocal microscopy images more accurately than naïve stereological models and significantly increases the feature-based prediction of red blood cell types from F1 = 79.0% to F1 = 87.4%. Applied to 2D images containing spheroidal aggregates of densely grown human induced pluripotent stem cells, we find that SHAPR learns fundamental shape properties of cell nuclei and allows for prediction-based morphometry. Reducing imaging time and data storage, SHAPR will help to optimize and up-scale image-based high-throughput applications for biomedicine.

Investigating phenotypic heterogeneity can help to better understand and manage microbial communities. However, characterizing phenotypic heterogeneity remains a challenge, as there is no standardized analysis framework. Several optical tools are available, such as flow cytometry and Raman spectroscopy, which describe optical properties of the individual cell. In this work, we compare Raman spectroscopy and flow cytometry to study phenotypic heterogeneity in bacterial populations. The growth stages of three replicate Escherichia coli populations were characterized using both technologies. Our findings show that flow cytometry detects and quantifies shifts in phenotypic heterogeneity at the population level due to its high-throughput nature. Raman spectroscopy, on the other hand, offers a much higher resolution at the single-cell level (i.e., more biochemical information is recorded). Therefore, it can identify distinct phenotypic populations when coupled with analyses tailored toward single-cell data. In addition, it provides information about biomolecules that are present, which can be linked to cell functionality. We propose a computational workflow to distinguish between bacterial phenotypic populations using Raman spectroscopy and validated this approach with an external data set. We recommend using flow cytometry to quantify phenotypic heterogeneity at the population level, and Raman spectroscopy to perform a more in-depth analysis of heterogeneity at the single-cell level. (c) 2019 International Society for Advancement of Cytometry

Multiple organ failure syndrome (MOFS) is a hallmark of sepsis. The continued dysfunction of microvascular perfusion has been implicated as the inciting factor for this. Multiple etiopathological factors are involved in producing this disequilibrium in the demand, supply, and extraction of oxygen as a result of derecruitment of microcirculation. This is further complicated by mitochondrial dysfunction in the form of the inhibition of mitochondrial respiratory chain of enzymes, leading to difficulty in extraction of the oxygen at the cellular level. Eventually, although hemodynamic stability of systemic/macrovascular circulation may have been achieved, the process of deficient and defective delivery of oxygen to the tissues goes on relentlessly. The indicators and monitoring of this process of impairment of oxygen delivery (DO2) have been discussed in this review. In addition, the review also encompasses various therapeutic modalities and their efficacy, based on the evidence. The latest guidelines regarding optimizing the DO2 in sepsis are also included here.

Chronic toxicity studies are an integral part of the overall toxicological evaluation of drug candidates. Biochemical studies of blood and urine serve to describe the general picture of the animal’s condition and to trace the dynamics of laboratory markers of organ damage after repeated administration of the drug — both for a group of animals, as well as individually. Pathomorphological studies of organs and tissues make it possible to understand the nature of damage at the cellular level. A systemic analysis of the data obtained allows for accurate identification of the body systems that are most susceptible to the toxic effect of the drug, as well as for assessment of the degree of impact and the reversibility of effects. Considering the great importance of the obtained pharmacological information, it is necessary to pay special attention to proper methodological implementation of all the stages of preparation and performance of the mentioned studies, as well as to proper analysis and interpretation of the data. The aim of the paper was to summarise the methodology of biochemical and pathomorphological studies based on the experience obtained in the Drug Toxicology Laboratory of the Federal State Budgetary Institute «Zakusov Institute of Pharmacology» when conducting chronic toxicity studies.

Optical trapping (synonymous with optical tweezers) has become a core biophysical technique widely used for interrogating fundamental biological processes on size scales ranging from the single-molecule to the cellular level. Recent advances in nanotechnology have led to the development of 'nanophotonic tweezers,' an exciting new class of 'on-chip' optical traps. Here, we describe how nanophotonic tweezers are making optical trap technology more broadly accessible and bringing unique biosensing and manipulation capabilities to biological applications of optical trapping.

Coral reefs are one of the most important ecosystems in the world but least understood from a cellular level. In this issue of Cell, Levy et al. unravel the single-cell gene expression of the coral holobiont and open the doors to better understand the novel diversity of cell types.