Author Correction: Mechanosensing through immunoreceptors.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Mechanosensing through immunoreceptors.

The immune response is orchestrated by a variety of immune cells. The function of each cell is determined by the collective signals from various immunoreceptors, whose expression and activity depend on the developmental stages of the cell and its environmental context. Recent studies have highlighted the presence of mechanical force on several immunoreceptor-ligand pairs and the important role of force in regulating their interaction and function. In this Perspective, we use the T cell antigen receptor as an example with which to review the current understanding of the mechanosensing properties of immunoreceptors. We discuss the types of forces that immunoreceptors may encounter and the effects of force on ligand bonding, conformational change and the triggering of immunoreceptors, as well as the effects of force on the downstream signal transduction, cell-fate decisions and effector function of immune cells.

A TCR mechanotransduction signaling loop induces negative selection in the thymus.

The T cell antigen receptor (TCR) expressed on thymocytes interacts with self-peptide major histocompatibility complex (pMHC) ligands to signal apoptosis or survival. Here, we found that negative-selection ligands induced thymocytes to exert forces on the TCR and the co-receptor CD8 and formed cooperative TCR-pMHC-CD8 trimolecular 'catch bonds', whereas positive-selection ligands induced less sustained thymocyte forces on TCR and CD8 and formed shorter-lived, independent TCR-pMHC and pMHC-CD8 bimolecular 'slip bonds'. Catch bonds were not intrinsic to either the TCR-pMHC or the pMHC-CD8 arm of the trans (cross-junctional) heterodimer but resulted from coupling of the extracellular pMHC-CD8 interaction to the intracellular interaction of CD8 with TCR-CD3 via associated kinases to form a cis (lateral) heterodimer capable of inside-out signaling. We suggest that the coupled trans-cis heterodimeric interactions form a mechanotransduction loop that reinforces negative-selection signaling that is distinct from positive-selection signaling in the thymus.

Characterization Techniques of Polymer Aging: From Beginning to End.

Polymers have been widely applied in various fields in the daily routines and the manufacturing. Despite the awareness of the aggressive and inevitable aging for the polymers, it still remains a challenge to choose an appropriate characterization strategy for evaluating the aging behaviors. The difficulties lie in the fact that the polymer features from the different aging stages require different characterization methods. In this review, we present an overview of the characterization strategies preferable for the initial, accelerated, and late stages during polymer aging. The optimum strategies have been discussed to characterize the generation of radicals, variation of functional groups, substantial chain scission, formation of low-molecular products, and deterioration in the polymers' macro-performances. In view of the advantages and the limitations of these characterization techniques, their utilization in a strategic approach is considered. In addition, we highlight the structure-property relationship for the aged polymers and provide available guidance for lifetime prediction. This review could allow the readers to be knowledgeable of the features for the polymers in the different aging stages and provide access to choose the optimum characterization techniques. We believe that this review will attract the communities dedicated to materials science and chemistry.

The kinetics of E-selectin- and P-selectin-induced intermediate activation of integrin αLß2 on neutrophils.

Selectins and integrins are key players in the adhesion and signaling cascade that recruits leukocytes to inflamed tissues. Selectin binding induces ß2 integrin binding to slow leukocyte rolling. Here, a micropipette was used to characterize neutrophil adhesion to E-selectin and intercellular adhesion molecule-1 (ICAM-1) at room temperature. The time-dependent adhesion frequency displayed two-stage kinetics, with an E-selectin-mediated fast increase to a low plateau followed by a slow increase to a high plateau mediated by intermediate-affinity binding of integrin αLß2 to ICAM-1. The αLß2 activation required more than 5 s contact to E-selectin and spleen tyrosine kinase (Syk) activity. A multi-zone channel was used to analyze αLß2 activation by P-selectin in separate zones of receptors or antibodies, finding an inverse relationship between the rolling velocity on ICAM-1 and P-selectin dose, and a P-selectin dose-dependent change from bent to extended conformations with a closed headpiece that was faster at 37°C than at room temperature. Activation of αLß2 exhibited different levels of cooperativity and persistent times depending on the strength and duration of selectin stimulation. These results define the precise timing and kinetics of intermediate activation of αLß2 by E- and P-selectins.

Superoxide flashes in single mitochondria.

In quiescent cells, mitochondria are the primary source of reactive oxygen species (ROS), which are generated by leakiness of the electron transport chain (ETC). High levels of ROS can trigger cell death, whereas lower levels drive diverse and important cellular functions. We show here by employing a newly developed mitochondrial matrix-targeted superoxide indicator, that individual mitochondria undergo spontaneous bursts of superoxide generation, termed "superoxide flashes." Superoxide flashes occur randomly in space and time, exhibit all-or-none properties, and provide a vital source of superoxide production across many different cell types. Individual flashes are triggered by transient openings of the mitochondrial permeability transition pore stimulating superoxide production by the ETC. Furthermore, we observe a flurry of superoxide flash activity during reoxygenation of cardiomyocytes after hypoxia, which is inhibited by the cardioprotective compound adenosine. We propose that superoxide flashes could serve as a valuable biomarker for a wide variety of oxidative stress-related diseases.

Study on the Thermal Stabilizing Process of Layered Double Hydroxides in PVC Resin.

Poly(vinyl chloride) (PVC) is widely used in various fields and requires the use of thermal stabilizers to enhance its thermal stability during processing because of its poor thermal stability. Layered double hydroxides (LDHs) are widely considered to be one kind of highly efficient and environmentally friendly PVC thermal stabilizer. To investigate the thermal stabilizing process of layered double hydroxides (LDHs) in PVC resin, PVC and MgAl-LDHs powders with different interlayer anions (CO32-, Cl-, and NO3-) were physically mixed and aged at 180 °C. The structure of LDHs at different aging times was studied using XRD, SEM, and FT-IR. The results show that the thermal stabilizing process of LDHs on PVC mainly has three stages. In the first stage, the layers of LDHs undergo a reaction with HCl, which is released during the thermal decomposition of PVC. Subsequently, the ion exchange process occurs between Cl- and interlayer CO32-, resulting in the formation of MgAl-Cl-LDHs. Finally, the layers of MgAl-Cl-LDHs react with HCl slowly. During the thermal stabilizing process of MgAl-Cl-LDHs, the peak intensity of XRD reduces slightly, and no new XRD peak emerges. It indicates that only the first step happens for MgAl-Cl-LDHs. The TG-DTA analysis of LDHs indicates that the interaction of LDHs with different interlayer anions has the following order: NO3- < CO32- < Cl-, according to the early coloring in the thermal aging test of PVC composites. The results of the thermal aging tests suggest that LDHs with a weak interaction between interlayer anions and layers can enhance the early stability of PVC significantly. Furthermore, the thermal aging test demonstrates that LDHs with high HCl absorption capacities exhibit superior long-term stabilizing effects on PVC resin. This finding provides a valuable hint for designing an LDHs/PVC resin with a novel structure and excellent thermal stability.

Frontal-Temporal Triangle Area Hair for Eyebrow Restoration in Asians: A Comparative Study With Periauricular and Occipital Hair.

BACKGROUND: The frontal-temporal triangle area (FTTA) hair has a slow growth rate and thin caliber, which are similar to those of eyebrow hair. However, a comparison of cosmetic outcomes between FTTA and other scalp hair grafts in eyebrow transplantation has not been performed. OBJECTIVE: To compare the cosmetic outcomes of FTTA hair and periauricular and occipital area (POA) hair in eyebrow restoration. METHODS: A retrospective analysis of 155 patients with FTTA or POA hair transplants was performed. Comparative variables included patient characteristics, hair density, diameter, percentage of one-hair follicular units (FUs), number of transplanted FUs, harvesting time, transection rate, hair survival rate, frequency of eyebrow trimming, and patient satisfaction. RESULTS: There was a significant difference in hair density, diameter, percentage of one-hair FUs, and harvesting time between the FTTA and POA hair transplants. The FTTA hair grew significantly slower than the POA hair did. The patients in the FTTA group trimmed their postoperative eyebrows at a significantly longer interval than those in the POA group. The percentage of patients who were very satisfied with the surgery results was higher in the FTTA group. CONCLUSION: The FTTA hair grafts can provide aesthetically pleasing cosmetic results in eyebrow restoration.

Effect of De-epithelialization on Graft Survival Rate After Follicular Unit Extraction.

BACKGROUND: To achieve a natural postoperative appearance, hair grafts are often de-epithelialized from the epidermis during follicular unit extraction (FUE). However, the effect of de-epithelialization on the survival rate of transplanted hair follicles (HFs) has not been investigated. OBJECTIVE: To investigate the effect of de-epithelialization on the survival rate of transplanted HFs. METHODS: A total of 64 male patients with androgenetic alopecia were included in this study. They were randomly divided into de-epithelialization and control groups. Organ culture was performed to assess the elongation of hair shaft and the percentage of anagen HFs in both groups. Patients were followed up postoperatively to evaluate complications, postoperative shedding, survival rates, and satisfaction. RESULTS: No significant difference in hair shaft elongation and percentage of anagen HFs was observed between both groups. The immediate postoperative satisfaction in the control group was much lower than that in the de-epithelialization group (71.25% and 100%, respectively). No significant differences in shedding rate, graft survival rate, and complications were noticed between both groups. CONCLUSION: Follicular de-epithelialization does not affect the survival rate of graft in FUE. Based on these data, de-epithelialization may improve immediate postoperative appearance and lead to a more pleasing cosmetic outcome.

Novel Fluorescence Method for Determination of Spatial Interparticle Distance in Polymer Nanocomposites.

Interparticle distance (ID) is generally used for spatial evaluation on the dispersion of nanofillers in polymer nanocomposites (PNCs) in order to gain in-depth insight into fundamental understanding of reinforcement and structure-property correlation. However, currently available methods mainly rely on two-dimensional observation technologies or simulation methods. Herein, using layered double hydroxides (LDHs) in low-density polyethylene (LDPE) as a model matrix, we developed a novel spatial ID determination method through a post labeling fluorescent imaging technique. The spatial ID of LDH nanofillers was achieved by a MATLAB program based on the 3D coordinates of LDH nanofillers in PNCs. The spatial ID data indicated the varied dispersion states of LDH nanofillers from "random", "even", to "clustered" in PNCs. More importantly, the so-called exceptional performances of PNCs at high loading of LDH nanofillers can be reasonably explained in combination with the mechanical studies. This proposed approach could undoubtedly provide valuable information in elucidating the structure-property correlation in PNCs. We believe that this work would be a guide to design advanced PNC materials.

In situ and in silico kinetic analyses of programmed cell death-1 (PD-1) receptor, programmed cell death ligands, and B7-1 protein interaction network.

Programmed cell death-1 (PD-1) is an inhibitory receptor with an essential role in maintaining peripheral tolerance and is among the most promising immunotherapeutic targets for treating cancer, autoimmunity, and infectious diseases. A complete understanding of the consequences of PD-1 engagement by its ligands, PD-L1 and PD-L2, and of PD-L1 binding to B7-1 requires quantitative analysis of their interactions at the cell surface. We present here the first complete in situ kinetic analysis of the PD-1/PD-ligands/B7-1 system. Consistent with previous solution measurements, we observed higher in situ affinities for human (h) than murine (m) PD-1 interactions, stronger binding of hPD-1 to hPD-L2 than hPD-L1, and comparable binding of mPD-1 to both ligands. However, in contrast to the relatively weak solution affinities, the in situ affinities of PD-1 are as high as those of the T cell receptor for agonist pMHC and of LFA-1 (lymphocyte function-associated antigen 1) for ICAM-1 (intercellular adhesion molecule 1) but significantly lower than that of the B7-1/CTLA-4 interaction, suggesting a distinct basis for PD-1- versus CTLA-4-mediated inhibition. Notably, the in situ interactions of PD-1 are much stronger than that of B7-1 with PD-L1. Overall, the in situ affinity ranking greatly depends on the on-rate instead of the off-rate. In silico simulations predict that PD-1/PD-L1 interactions dominate at interfaces between activated T cells and mature dendritic cells and that these interactions will be highly sensitive to the dynamics of PD-L1 and PD-L2 expression. Our results provide a kinetic framework for better understanding inhibitory PD-1 activity in health and disease.

Calibration of the degree of linear polarization measurements of the polarized Sun-sky radiometer based on the POLBOX system.

Polarization observation of sky radiation is the frontier approach to improve the remote sensing of atmospheric components, e.g., aerosol and clouds. The polarization calibration of the ground-based Sun-sky radiometer is the basis for obtaining accurate degree of linear polarization (DOLP) measurement. In this paper, a DOLP calibration method based on a laboratory polarized light source (POLBOX) is introduced in detail. Combined with the CE318-DP Sun-sky polarized radiometer, a calibration scheme for DOLP measurement is established for the spectral range of 440-1640 nm. Based on the calibration results of the Sun-sky radiometer observation network, the polarization calibration coefficient and the DOLP calibration residual are analyzed statistically. The results show that the DOLP residual of the calibration scheme is about 0.0012, and thus it can be estimated that the final DOLP calibration accuracy of this method is about 0.005. Finally, it is verified that the accuracy of the calibration results is in accordance with the expected results by comparing the simulated DOLP with the vector radiative transfer calculations.

Simple transfer calibration method for a Cimel Sun-Moon photometer: calculating lunar calibration coefficients from Sun calibration constants.

The Cimel new technologies allow both daytime and nighttime aerosol optical depth (AOD) measurements. Although the daytime AOD calibration protocols are well established, accurate and simple nighttime calibration is still a challenging task. Standard lunar-Langley and intercomparison calibration methods both require specific conditions in terms of atmospheric stability and site condition. Additionally, the lunar irradiance model also has some known limits on its uncertainty. This paper presents a simple calibration method that transfers the direct-Sun calibration constant, V0,Sun, to the lunar irradiance calibration coefficient, CMoon. Our approach is a pure calculation method, independent of site limits, e.g., Moon phase. The method is also not affected by the lunar irradiance model limitations, which is the largest error source of traditional calibration methods. Besides, this new transfer calibration approach is easy to use in the field since CMoon can be obtained directly once V0,Sun is known. Error analysis suggests that the average uncertainty of CMoon over the 440-1640 nm bands obtained with the transfer method is 2.4%-2.8%, depending on the V0,Sun approach (Langley or intercomparison), which is comparable with that of lunar-Langley approach, theoretically. In this paper, the Sun-Moon transfer and the Langley methods are compared based on site measurements in Beijing, and the day-night measurement continuity and performance are analyzed.

Autophagy induces hair follicle stem cell activation and hair follicle regeneration by regulating glycolysis.

BACKGROUND: Hair follicle stem cells (HFSCs) typically remain quiescent and are activated only during the transition from telogen to anagen to ensure that the hair follicle enters a new cycle. The metabolic behavior of stem cells in tissues is regulated by macroautophagy/autophagy, and changes in HFSC metabolism directly affect their activation and maintenance. However, the role of autophagy in the regulation of HFSC metabolism and function remains unclear. METHODS: Back skin samples were obtained from mice at different hair follicle cycle stages, and immunofluorescence staining was used to monitor autophagy in HFSCs. Mouse and human hair follicles were treated with rapamycin (Rapa, an autophagy activator) or 3-methyladenine (3-MA, an autophagy inhibitor). The effects of autophagy on the hair follicle cycle and HFSC were investigated by imaging, cell proliferation staining, and HFSC-specific marker staining. The influence and mechanism of autophagy on HFSC metabolism were explored using RNA sequencing, real-time polymerase chain reaction, immunohistochemical staining, and detection of lactate and glucose concentrations. Finally, the influence of autophagy-induced glycolysis on HFSC and the hair follicle cycle was verified by stem cell characteristics and in vivo functional experiments. RESULTS: Autophagy in HFSC was highest during the transition from telogen to anagen. Inhibiting autophagy with 3-MA led to early entry into catagen and prolonged telogen, whereas Rapa promoted autophagy and hair growth. Autophagy activated HFSC by increasing the expression and activity of HFSC lactate dehydrogenase (Ldha), thereby transforming HFSC metabolism into glycolysis. Inhibition of Ldha expression counteracted the effects of autophagy. CONCLUSIONS: Autophagy activated HFSC by promoting the transition from HFSC metabolism to glycolysis, ultimately initiating the hair follicle cycle and promoting hair growth.

Light-field flow cytometry for high-resolution, volumetric and multiparametric 3D single-cell analysis.

Imaging flow cytometry (IFC) combines flow cytometry and fluorescence microscopy to enable high-throughput, multiparametric single-cell analysis with rich spatial details. However, current IFC techniques remain limited in their ability to reveal subcellular information with a high 3D resolution, throughput, sensitivity, and instrumental simplicity. In this study, we introduce a light-field flow cytometer (LFC), an IFC system capable of high-content, single-shot, and multi-color acquisition of up to 5,750 cells per second with a near-diffraction-limited resolution of 400-600 nm in all three dimensions. The LFC system integrates optical, microfluidic, and computational strategies to facilitate the volumetric visualization of various 3D subcellular characteristics through convenient access to commonly used epi-fluorescence platforms. We demonstrate the effectiveness of LFC in assaying, analyzing, and enumerating intricate subcellular morphology, function, and heterogeneity using various phantoms and biological specimens. The advancement offered by the LFC system presents a promising methodological pathway for broad cell biological and translational discoveries, with the potential for widespread adoption in biomedical research.

Method to intercalibrate sunphotometer constants using an integrating sphere as a light source in the laboratory.

A calibration method is introduced to transfer calibration constants from the reference to secondary sunphotometers using a laboratory integrating sphere as a light source, instead of the traditional transferring approach performed at specific calibration sites based on sunlight. The viewing solid angle and spectral response effects of the photometer are taken into account in the transfer, and thus the method can be applied to different types of sunphotometers widely used in the field of atmospheric observation. A laboratory experiment is performed to illustrate this approach for four types of CIMEL CE318 sunphotometers belonging to the aerosol robotic network (AERONET). The laboratory calibration method shows an average difference of 1.4% from the AERONET operational calibration results, while a detailed error analysis suggests that the uncertainty agrees with the estimation and could be further improved. Using this laboratory calibration approach is expected to avoid weather influences and decrease data interruption due to operationally required periodic calibration operations. It also provides a basis for establishing a network including different sunphotometers for worldwide aerosol measurements, based on a single standard calibration reference.

[Improving accumulation-mode fraction based on spectral aerosol optical depth in Beijing].

In the present study, the authors put forward a parameterization method of correcting fine-mode Angstrom index with aerosol optical depth and volume distribution of aerosol derived from AERONET in Beijing over 2011. The parameterization method is coupled with aerosol optical depth spectral deconvolution algorithm to improve the accuracy of accumulation-mode fraction. The errors of estimated AMF are derived from underestimate of fine-mode Angstrom index errors. We calculate and simulate the extreme values of fine-mode Angstrom index, getting constraint conditions, and then establish the extreme values correction method. Results from sensitivity test suggest that extreme values of fine-mode Angstrom index are constrained in the reasonable range. Finemode Angstrom index and AMF are sensitive to normalized volume distribution of aerosol, ranging from 0.662 to 2.849 and from 0.08 to 0.84 due to different distribution. Mean deviation of accumulation-mode fraction is reduced from 0.072 to 0.044, and the difference is 38.89%, especially in winter and summer. Improving computational accuracy of accumulation-mode fraction can enhance that of anthropogenic aerosol optical thickness, and it has important significance of anthropogenic aerosol direct radiation force estimation and environmental quality assessment.

Design of mechanical-robust phosphorescence materials through covalent click reaction.

It remains a great challenge to engineer materials with strong and stable interactions for the simultaneously mechanical-robust and room temperature phosphorescence-efficient materials. In this work, we demonstrate a covalent cross-linking strategy to engineer mechanical-robust room temperature phosphorescence materials through the B-O click reaction between chromophores, polyvinyl alcohol matrix and inorganic layered double hydroxide nanosheets. Through the covalent cross-linkage between the organic polyvinyl alcohol and inorganic layered double hydroxide, a polymeric composite with ultralong lifetime up to 1.45 s is acquired based on the inhibited non-radiative transition of chromophores. Simultaneously, decent mechanical strength of 97.9 MPa can be realized for the composite materials due to the dissipated loading stress through the covalent-bond-accommodated interfacial interaction. These cross-linked composites also exhibit flexibility, processability, scalability and phosphorescence responses towards the mechanical deformation. It is anticipated that the proposed covalent click reaction could provide a platform for the design and modulation of composites with multi-functionality and long-term durability.

Addressing the Origin of Single-Atom-Activated Supports Monitored by Electrochemiluminescence.

Currently, much attention has been paid to the efforts to stabilize and regulate single atoms through supports to obtain decent electrocatalytic behaviors. However, little concern was given to the effect of single atoms on modulating the electronic structure of supports, despite the catalytic activities and large quantities of supports in the catalytic reactions. Herein, we have localized Ru single atoms onto two-dimensional layered double hydroxide (NiFe-LDH) and studied the role of Ru single atoms in adjusting the electronic structure of the NiFe-LDH support. Spin polarization of 3d electrons for Fe and electron redistribution in NiFe-LDH were effectively modulated through the interaction between Ru single atoms and NiFe-LDH. As a result, the luminol redox reaction and reactive oxygen revolution were simultaneously promoted by Ru single-atom-modulated NiFe-LDH, manifested as boosted electrochemiluminescence (ECL). Therefore, we have provided valid information to reveal the regulation effect of single atoms on the spin state and electronic structure of the supports. It is anticipated that our strategy may arouse wide interest in manipulating single-atom-modulated supports.