Mechanistic Insights into the Folding Mechanism of Region V in Ice-Binding Protein Secreted by Marinomonas primoryensis Revealed by Single-Molecule Force Spectroscopy.

The Gram-negative bacteria Marinomonas primoryensis secrete an ice-binding protein (MpIBP), which is a vital bacterial adhesin facilitating the adaptation and survival of the bacteria in the harsh Antarctic environment. The C-terminal region of MpIBP, known as region V (RV), is the first domain to be exported into the Ca2+-rich extracellular environment and acts as a folding nucleus for the entire adhesin. However, the mechanisms underlying the secretion and folding of RV remain poorly understood. Here, we used optical tweezers (OT) to investigate the secretion and folding mechanisms of RV at the single-molecule level. In the absence of Ca2+, apo-RV remains unstructured, while Ca2+-bound RV folds into a mechanically stable structure. The folding of RV could occur via the formation of an intermediate state. Even though this folding intermediate is "hidden" during the folding process of wild type RV in vitro, it likely forms in vivo and plays an important role in facilitating protein secretion. Additionally, our results revealed that the N-terminal part of the RV can significantly stabilize its C-terminal structure. Our study paves the way for further investigations into the structure and functions of MpIBP that help bacteria survive in challenging environments.

Microsphere probe: combining microsphere-assisted microscopy with AFM.

In recent years, microsphere-assisted microscopy (MAM) and atomic force microscope (AFM) have been rapidly developed to meet the measurement needs of microstructures. However, the positioning of microspheres, the inability of AFM to touch the underlying sample through the transparent insulating layer, and the challenge of AFM fast positioning limit their use in practical measurements. In this paper, we propose a method that combines MAM with AFM by adhering the microsphere to the cantilever. This method allows MAM and AFM to work in parallel, and their imaging positions can correspond with each other. We use this method to measure memory devices, and the results show that MAM and AFM yield complementary advantages. This approach provides a new tool for analyzing complex structures in devices and has potential for wide application.

Deep learning for precise axial localization of trapped microspheres in reflective optical systems.

High-precision axial localization measurement is an important part of micro-nanometer optical measurement, but there have been issues such as low calibration efficiency, poor accuracy, and cumbersome measurement, especially in reflected light illumination systems, where the lack of clarity of imaging details leads to the low accuracy of commonly used methods. Herein, we develop a trained residual neural network coupled with a convenient data acquisition strategy to address this challenge. Our method improves the axial localization precision of microspheres in both reflective illumination systems and transmission illumination systems. Using this new localization method, the reference position of the trapped microsphere can be extracted from the identification results, namely the "positioning point" among the experimental groups. This point relies on the unique signal characteristics of each sample measurement, eliminates systematic repeatability errors when performing identification across samples, and improves the localization precision of different samples. This method has been verified on both transmission and reflected illumination optical tweezers platforms. We will bring greater convenience to measurements in solution environments and will provide higher-order guarantees for force spectroscopy measurements in scenarios such as microsphere-based super-resolution microscopy and the surface mechanical properties of adherent flexible materials and cells.

Addressing the imaging limitations of a microsphere-assisted nanoscope.

In the past decade, microsphere-assisted nanoscopy has been developed rapidly to overcome the diffraction limit. However, due to the limited size and high surface curvature of microspheres, the magnified imaging still suffers from problems like limited view scope, imaging distortion, and low contrast. In this paper, we specialize in the imaging mechanism of microspheres and find irradiance as the key factor for microsphere imaging quality. Utilizing a modified optical tweezer system, we achieve precise manipulation of microspheres and further propose a high-quality large-field magnified imaging scheme. The results show that the imaging area of 5 µm microspheres can reach 16×12 µm2 with the minimum identifiable feature of 137 nm. This scheme provides a new solution for extending the measuring scope of microsphere-assisted nanoscope, and will certainly promote the application of this technology in practice.

Simulation and Experiment of the Trapping Trajectory for Janus Particles in Linearly Polarized Optical Traps.

The highly focused laser beam is capable of confining micro-sized particle in its focus. This is widely known as optical trapping. The Janus particle is composed of two hemispheres with different refractive indexes. In a linearly polarized optical trap, the Janus particle tends to align itself to an orientation where the interface of the two hemispheres is parallel to the laser propagation as well as the polarization direction. This enables a controllable approach that rotates the trapped particle with fine accuracy and could be used in partial measurement. However, due to the complexity of the interaction of the optical field and refractive index distribution, the trapping trajectory of the Janus particle in the linearly polarized optical trap is still uncovered. In this paper, we focus on the dynamic trapping process and the steady position and orientation of the Janus particle in the optical trap from both simulation and experimental aspects. The trapping process recorded by a high speed camera coincides with the simulation result calculated using the T-matrix model, which not only reveals the trapping trajectory, but also provides a practical simulation solution for more complicated structures and trapping motions.

Bag-1L Protects against Cell Apoptosis in an In Vitro Model of Lung Ischemia-Reperfusion Injury through the C-Terminal "Bag" Domain.

Bcl-2-associated athanogene 1 (Bag-1) is a multifunctional and antiapoptotic protein that binds to the antiapoptosis regulator Bcl-2 and promotes cell survival. To investigate the protective function of Bag-1, we examined the effects of Bag-1L, one isoform of Bag-1, in an in vitro cell culture model of lung ischemia-reperfusion injury (LIRI) generated by treatment of A549 cells with hypoxia/reoxygenation. Overexpression of full-length Bag-1L increased the viability of A549 cells and reduced cell apoptosis in response to 6 h of hypoxia/reoxygenation treatment. Furthermore, Bag-1L overexpression enhanced the heat shock protein 70 (HSP70) and Bcl-2 protein levels, increased the phosphorylation of AKT, decreased Bax and cleaved caspase-3 levels, and was able to overcome cell cycle arrest. These effects were not observed in A549 cells overexpressing a truncated form of Bag-1L lacking the "Bag domain," denoted Bag-1L△C. The "Bag domain" is the C-terminal 47 amino acids. Taken together, the results of this study suggest that Bag-1L overexpression can protect against oxidative stress and apoptosis in an in vitro LIRI model, with a dependence on the Bag domain.

The formation principle of micro-droplets induced by using optical tweezers.

Utilizing droplets as micro-tools has become a valuable method in biology and chemistry. In previous work, we have demonstrated a novel droplet generation-manipulation method in a conventional optical tweezer system. Here, a further study of the droplet composition and its formation principle is performed. First, it is proved through Raman spectra that the principal component of the droplets is HPO4 2- solution. Considering that the generated droplet size is at the µm level, we have adopted a variety of methods in experiments to reduce external interference. Second, using a confocal microscopic video camera, the growth process of the droplet is completely recorded in a common glass-based chamber. The finite element simulations help us to further understand that the droplet generation process using optical tweezers can be divided into two stages: "capture" caused by optical force field and "aggregation" induced by a photothermal phenomenon and thermal acceleration. Through these studies, the nature of the optical tweezer-generated droplets is revealed. As a general principle for the droplet generation, this method will provide inspiration and prospects in the fields of microfluidics and biophysics-chemistry.

Cooperative Adaptive Cruise Control and exhaust emission evaluation under heterogeneous connected vehicle network environment in urban city.

With the development of information communication and artificial intelligence, the ICV (intelligent connected vehicle) will inevitably play an important part in future urban transport system. In this paper, we study the car following behaviour under the heterogeneous ICV environment. The time to receive information varies from vehicle to vehicle, since the manual vehicles and autonomous vehicles co-exist on the road. By introducing time-varying lags function, a new car following model is proposed, and the cooperative control strategy of this model is studied. Based on Lyapunov function theory and linear matrix inequality (LMI) approach, the sufficient condition that the existence of the feedback controller is given, which makes the closed-loop system asymptotically stable under mixed traffic flow environment. That is to say, traffic congestion phenomenon under heterogeneous traffic flow can be effectively suppressed, and the feedback controller gain matrix can be obtained via solving linear matrix inequality. Finally, by simulation the method is verified effective in alleviating traffic congestions and reducing fuel consumption and exhaust emissions. It could be a useful reference to Cooperative Vehicle Infrastructure System and Smart City.

Safety and complications of medical thoracoscopy in the management of pleural diseases.

BACKGROUND: Medical thoracoscopy is considered an overall safe procedure, whereas numbers of studies focus on complications of diagnostic thoracoscopy and talc poudrage pleurodesis. We conduct this study to evaluate the safety of medical thoracoscopy in the management of pleural diseases and to compare complications in different therapeutic thoracoscopic procedures. METHODS: A retrospective study was performed in 1926 patients, 662 of whom underwent medical thoracoscopy for diagnosis and 1264 of whom for therapeutic interventions of pleural diseases. Data on complications were obtained from the patients, notes on computer system, laboratory and radiographic findings. Chi-square test was performed to compare categorical variables and Fisher's exact test was used for small samples. RESULTS: The mean age was 51 ± 8.4 (range 21-86) years and 1117 (58%) were males. Diagnostic procedure was taken in 662 (34.4%) patients, whereas therapeutic procedure was taken in 1264 (65.6%) patients. Malignant histology was reported in 860 (44.6%) and 986 (51.2%) revealed benign pleural diseases. Eighty patients (4.2%) were not definitely diagnosed and they were considered as unidentified pleural effusion. One patient died during the creation of artificial pneumothorax, and the causes of death were supposed as air embolism or an inhibition of phrenic motoneurons and circulatory system. Complication of lung laceration was found in six patients (0.3%) and reexpansion pulmonary edema was observed in two patients (0.1%). Higher incidence of prolonged air leak was observed in bulla electrocoagulation group, in comparison with pleurodesis group. Moreover, pain and fever were the most frequently complications in pleurodesis group and cutaneous infection in entry site was the most frequently reported complication in pleural decortication of empyema group. CONCLUSIONS: Medical thoracoscopy is generally a safe and effective method, not only in the diagnosis of undiagnosed pleural effusions, but also in the management of pleural diseases. Mastering medical thoracoscopy well, improving patient management after the procedure and attempts to reduce the occurrence of post-procedural complications are the targets that physicians are supposed to achieve in the future.

Erythromycin poudrage versus erythromycin slurry in the treatment of refractory spontaneous pneumothorax.

BACKGROUND: Refractory (recurrent or persistent) spontaneous pneumothorax with high recurrence rates required treatment either by continuous chest drainage or interventional approaches. Pleurodesis by sclerosing agents has become a significant therapy in the treatment of refractory spontaneous pneumothorax (RSP) on account of its high efficiency and safety. However, the efficacy, safety and appropriate mode of administration of intrapleural erythromycin for pleurodesis have not yet been realized in the treatment of RSP. METHODS: The trial was performed to compare thoracoscopic erythromycin poudrage with erythromycin slurry via a chest tube for patients with documented RSP. Fifty-seven patients with RSP were enrolled in this study with 30 patients for erythromycin poudrage and 27 patients for erythromycin slurry. Response to pleurodesis, complications and recurrences were recorded. Continuous variables were compared with t-test. Chi-square test was performed to compare categorical variables and Fisher's exact test was used for small samples. RESULTS: Twenty-four patients in the erythromycin poudrage group (80%) and sixteen in the erythromycin slurry (ES) group (59.26%) had an immediately successful pleurodesis within 5 days (P=0.087). Patients in erythromycin poudrage had shorter duration of postprocedural chest tube drainage (6.23±3.04 days) than patients in ES (10.67±9.81 days) (P=0.032). During the follow-up, there was no significant statistical difference in recurrence rates between the two groups. Common adverse reactions included fever and chest pain with no significant difference between the two groups. CONCLUSIONS: Erythromycin is an effective and safe sclerosing agent for pleurodesis in management of RSP. Both methods are safe but erythromycin poudrage is more effective than ES.

Core-shell coaxially structured triboelectric nanogenerator for energy harvesting and motion sensing.

Converting sustainable human motion energy into electric energy has become an urgent task for the advancement of next-generation wearable and portable electronics. Herein, a core-shell coaxially structured triboelectric nanogenerator (CSTN) was fabricated by inserting an inner hollow circular tube into an outer hollow circular tube, and a gasbag is constructed within the space between the inner and outer tubes. Both Ni-coated polyester conductive textile and the conductive silicone rubber were used as effective electrode materials. The CSTN has excellent properties, including flexibility, light weight, sustainability and biological compatibility due to its unique structural design and materials selection. The CSTN can convert various forms of human motion energy, such as pressing, bending and twisting motion, into electric energy. A high short-circuit current of 11 µA and an open-circuit voltage of 380 V can be obtained from a CSTN with a length of 6 cm, corresponding to a high peak power of 1.638 mW at a load resistance of about 10 MΩ. When six such CSTNs are connected in parallel and placed under shoes, the electric energy output by normal walking can light up 60 LEDs connected serially and power up a competition-timer. The device can also sense different bending angles or twisting angles according to its signal outputs under different deformation angles. This study indicates the promising application prospects of the CSTN for next-generation devices, including self-powered illuminating devices, portable electronics, body motion sensing and health monitoring.

Influence of orally fed a select mixture of Bacillus probiotics on intestinal T-cell migration in weaned MUC4 resistant pigs following Escherichia coli challenge.

Efficient strategies for treating enteritis caused by F4(+) enterotoxigenic Escherichia coli (ETEC)/verocytotoxigenic Escherichia coli (VTEC)/enteropathogenic E. coli (EPEC) in mucin 4 resistant (MUC4 RR; supposed to be F4ab/ac receptor-negative [F4ab/acR(-)]) pigs remain elusive. A low (3.9 × 10(8) CFU/day) or high (7.8 × 10(8) CFU/day) dose of Bacillus licheniformis and Bacillus subtilis spore mixture (BLS-mix) was orally administered to MUC4 RR piglets for 1 week before F4(+) ETEC/VTEC/EPEC challenge. Orally fed BLS-mix upregulated the expression of TLR4, NOD2, iNOS, IL-8, and IL-22 mRNAs in the small intestine of pigs challenged with E. coli. Expression of chemokine CCL28 and its receptor CCR10 mRNAs was upregulated in the jejunum of pigs pretreated with high-dose BLS-mix. Low-dose BLS-mix pretreatment induced an increase in the proportion of peripheral blood CD4(-)CD8(-) T-cell subpopulations and high-dose BLS-mix induced the expansion of CD4(-)CD8(-) T cells in the inflamed intestine. Immunostaining revealed that considerable IL-7Rα-expressing cells accumulated at the lamina propria of the inflamed intestines after E. coli challenge, even in pigs pretreated with either low- or high-dose BLS-mix, although Western blot analysis of IL-7Rα expression in the intestinal mucosa did not show any change. Our data indicate that oral administration of the probiotic BLS-mix partially ameliorates E. coli-induced enteritis through facilitating upregulation of intestinal IL-22 and IκBα expression, and preventing loss of intestinal epithelial barrier integrity via elevating ZO-1 expression. However, IL-22 also elicits an inflammatory response in inflamed intestines as a result of infection with enteropathogenic bacteria.