Cowpea lipid transfer protein 1 regulates plant defense by inhibiting the cysteine protease of cowpea mosaic virus.

Many virus genomes encode proteases that facilitate infection. The molecular mechanism of plant recognition of viral proteases is largely unexplored. Using the system of Vigna unguiculata and cowpea mosaic virus (CPMV), we identified a cowpea lipid transfer protein (LTP1) which interacts with CPMV-encoded 24KPro, a cysteine protease, but not with the enzymatically inactive mutant 24KPro(C166A). Biochemical assays showed that LTP1 inhibited 24KPro proteolytic cleavage of the coat protein precursor large coat protein-small coat protein. Transient overexpression of LTP1 in cowpea reduced CPMV infection, whereas RNA interference-mediated LTP1 silencing increased CPMV accumulation in cowpea. LTP1 is mainly localized in the apoplast of uninfected plant cells, and after CPMV infection, most of the LTP1 is relocated to intracellular compartments, including chloroplast. Moreover, in stable LTP1-transgenic Nicotiana benthamiana plants, LTP1 repressed soybean mosaic virus (SMV) nuclear inclusion a protease activity, and accumulation of SMV was significantly reduced. We propose that cowpea LTP1 suppresses CPMV and SMV accumulation by directly inhibiting viral cysteine protease activity.

Mitophagy in plants: Emerging regulators of mitochondrial targeting for selective autophagy.

The degradation and turnover of mitochondria is fundamental to Eukaryotes and is a key homeostatic mechanism for maintaining functional mitochondrial populations. Autophagy is an important pathway by which mitochondria are degraded, involving their sequestration into membrane-bound autophagosomes and targeting to lytic endosomal compartments (the lysosome in animals, the vacuole in plants and yeast). Selective targeting of mitochondria for autophagy, also known as mitophagy, distinguishes mitochondria from other cell components for degradation and is necessary for the regulation of mitochondria-specific cell processes. In mammals and yeast, mitophagy has been well characterised and is regulated by numerous pathways with diverse and important functions in the regulation of cell homeostasis, metabolism and responses to specific stresses. In contrast, we are only just beginning to understand the importance and functions of mitophagy in plants, chiefly as the proteins that target mitochondria for autophagy in plants are only recently emerging. Here, we discuss the current progress of our understanding of mitophagy in plants, the importance of mitophagy for plant life and the regulatory autophagy proteins involved in mitochondrial degradation. In particular, we will discuss the recent emergence of mitophagy receptor proteins that selectively target mitochondria for autophagy, and discuss the missing links in our knowledge of mitophagy-regulatory proteins in plants compared to animals and yeast.

Hybrid kyphoplasty with short-versus intermediate- and long-segment pedicle screw fixations for the management of thoracolumbar burst fractures.

BACKGROUND: This study aimed to determine if the hybrid short-segment (HSS) technique is a good alternative to the intermediate-segment (IS) and long-segment (LS) techniques in pedicle screw fixations for acute thoracolumbar burst fractures (TLBFs). METHODS: In this retrospective evaluation, we examined 43 patients who underwent surgical treatments, including one- or two-level suprajacent (U) and infrajacent (L) pedicle screw fixations, for acute single-level TLBFs with neurological deficits between the T11 and L2 levels from July 2013 to December 2019. Among these patients, 15 individuals underwent HSS (U1L1), 12 received IS (U2L1), and 16 underwent LS (U2L2) fixations. Supplemental kyphoplasty of the fractured vertebral bodies was performed exclusively in the HSS group. Our analysis focused on assessing blood loss and surgical duration. Additionally, we compared postoperative thoracolumbar kyphotic degeneration using the data on Cobb angles on lateral radiographic images acquired at three time points (preoperatively, postoperative day 1, and follow-up). The end of follow-up was defined as the most recent postoperative radiographic image or implant complication occurrence. RESULTS: Blood loss and surgical duration were significantly lower in the HSS group than in the IS and LS groups. Additionally, the HSS group exhibited the lowest implant complication rate (2/15, 13.33%), followed by the LS (6/16, 37.5%) and IS (8/12, 66.7%) group. Implant complications occurred at a mean follow-up of 7.5 (range: 6-9), 9 (range: 5-23), and 7 (range: 1-21) months in the HSS, IS, and LS groups. Among these implant complications, revision surgeries were performed in two patients in the HSS group, two in the IS group, and one in the LS group. One patient treated by HSS with balloon kyphoplasty underwent reoperation because of symptomatic cement leakage. CONCLUSIONS: The HSS technique reduced intraoperative blood loss, surgical duration, and postoperative implant complications, indicating it is a good alternative to the IS and LS techniques for treating acute single-level TLBFs. This technique facilitates immediate kyphosis correction and successful maintenance of the corrected alignment within 1 year. Supplemental kyphoplasty with SpineJack® devices and high-viscosity bone cements for anterior reconstruction can potentially decrease the risk of cement leakage and related issues.

Predictors of depressive and anxiety symptoms among lesbian, gay and bisexual young adult individuals experiencing the COVID-19 pandemic: A four-year follow-up study.

BACKGROUND/PURPOSE: Depressive and anxiety symptoms were common among lesbian, gay, and bisexual (LGB) individuals during the COVID-19 pandemic. This 4-year follow-up study was conducted to investigate the predictors of depressive and anxiety symptoms in Taiwan's young adult LGB population. METHODS: Baseline data, including depressive and anxiety symptoms, demographic characteristics, sexual stigma, self-identity confusion, and family support were collected from 1000 LGB individuals. The participants' depressive and anxiety symptoms were reassessed 4 years after the baseline measurements. The predictive effects of the baseline factors on depressive and anxiety symptoms at follow-up were examined through linear regression analysis. RESULTS: Greater lack of identity, unconsolidated identity, sexual orientation microaggression, and lower perceived family function at baseline were significantly associated with more severe depressive and anxiety symptoms at follow-up. After adjustment for baseline depressive symptoms, being men, greater lack of identity, lower perceived family function, and more severe anxiety symptoms at baseline were significantly associated with more severe depressive symptoms at follow-up. After adjustment for baseline anxiety symptoms, greater unconsolidated identity and more severe depressive symptoms at baseline were significantly associated with more severe anxiety symptoms at follow-up. CONCLUSIONS: Intervention aimed at reducing depressive and anxiety symptoms in LGB individuals should be developed considering the predictors identified in this study.

Research on Vehicle Pose Detection Method Based on a Roadside Unit.

Vehicle pose detection plays a vital role in modern automotive technology, which can improve driving safety, enhance vehicle stability and provide important support for the development of autonomous driving technology. The current pose estimation methods have the problems of accumulation errors, large algorithm computing power, and expensive cost, so they cannot be widely used in intelligent connected vehicles. This paper proposes a vehicle pose detection method based on an RSU (Roadside Unit). First, the on-board GPS performs the positioning of the target vehicle and transmits the positioning information to the RSU through the UDP (User Data Protocol). Next, the RSU transmits a forward command to the OBU (On-board Unit) through the UDP. The OBU sends the command to the ECU (Electronic Control Unit) to control the vehicle forward. Then, the RSU detects and tracks the vehicle. The RSU takes pictures of two images before and after the movement and obtains the coordinates of the four angle points and the center point by image processing. The vehicle heading direction is determined by the moving direction of the center point of the front and rear two images. Finally, the RSU captures the vehicle images in real time, performs the process of tracking, rectangular fitting and pose calculation to obtain the pose information and transmits the information to the OBU to complete the whole process of vehicle pose detection and information transmission. Experiments show that the method can realize accurate and efficient detection of vehicle pose, meet the real-time requirements of vehicle pose detection, and can be widely used in intelligent vehicles.

Make It Shine: Labelling the ER for Light and Fluorescence Microscopy.

The ER is a highly dynamic network of tubules and membrane cisternae. Hence, imaging this organelle in its native and mobile state is of great importance. Here we describe methods of labelling the native plant ER using fluorescent proteins and lipid dyes as well as methods for immunolabelling on plant tissue.

Studying Autophagy and Senescence of Arabidopsis Stigmatic Cells.

The pistil is the most important organ for fertilization in flowering plants, and the stigmatic papilla cells are responsible for pollen acceptance and pollen tube germination. Arabidopsis plants possess dry stigmas exhibiting high selectivity for compatible pollen. When compatible pollens are recognized and accepted by stigmatic papilla cells, water and nutrients are then transported from the stigma to pollen grains through the secretory pathway. Here, we present light microscopy-based methods for investigating autophagy and senescence of stigmatic papilla cells. These methods include the assessment of viability of stigmatic papilla cells using dual staining with fluorescein diacetate/propidium iodide, as well as the examination of vacuolar-accumulated proteins during stigma senescence. These methods can be used to understand the functions of the stigma tissue from a subcellular perspective.

Studying Plant ER-PM Contact Site Localized Proteins Using Microscopy.

As in most eukaryotic cells, the plant endoplasmic reticulum (ER) network is physically linked to the plasma membrane (PM), forming ER-PM contact sites (EPCS). The protein complex required for maintaining the EPCS is composed of ER integral membrane proteins (e.g., VAP27, synaptotagmins), PM-associated proteins (e.g., NET3C), and the cytoskeleton. Here, we describe methods for studying EPCS structures and identifying possible EPCS-associated proteins. These include using artificially constructed reporters, GFP tagged protein expression followed by image analysis, and immunogold labelling at the ultrastructural level. In combination, these methods can be used to identify the location of putative EPCS proteins, which can aid in predicting their potential subcellular function.

Assays to Study Mitophagy in Plants.

Like most eukaryotic cells, mitophagy is essential in plant development and stress response. Several recent studies have revealed proteins that regulate this process, such as Friendly (FMT) and TraB family proteins (TRB), which are plant-unique mitophagy regulators so far. Here, we describe methods for studying mitophagy activity in plants through conventional microscopy and the use of loss-of-function mutants, such as using transgenic mitochondrial marker lines followed by image analysis, chemical inhibitor treatment, and plant phenotype studies. These methods can be used in combination to identify the putative mitophagy regulators and understand their functions in mitochondrial-related activities in plants.

Exo84c-regulated degradation is involved in the normal self-incompatible response in Brassicaceae.

The self-incompatibility system evolves in angiosperms to promote cross-pollination by rejecting self-pollination. Here, we show the involvement of Exo84c in the SI response of both Brassica napus and Arabidopsis. The expression of Exo84c is specifically elevated in stigma during the SI response. Knocking out Exo84c in B. napus and SI Arabidopsis partially breaks down the SI response. The SI response inhibits both the protein secretion in papillae and the recruitment of the exocyst complex to the pollen-pistil contact sites. Interestingly, these processes can be partially restored in exo84c SI Arabidopsis. After incompatible pollination, the turnover of the exocyst-labeled compartment is enhanced in papillae. However, this process is perturbed in exo84c SI Arabidopsis. Taken together, our results suggest that Exo84c regulates the exocyst complex vacuolar degradation during the SI response. This process is likely independent of the known SI pathway in Brassicaceae to secure the SI response.

A novel single-port robot for total gastrectomy to treat gastric cancer: A case report (with video).

Multiport robots are now widely used for total gastrectomy for gastric cancer, while there is almost a void of research on whether single-port (SP) robots can be used for total gastrectomy. Here, we report a case of a 75-year-old female patient who was diagnosed with gastric cardia adenocarcinoma by gastroscopy and underwent total gastrectomy assisted by the SHURUI SP robot. We successfully accomplished total gastrectomy and D2 lymph node dissection using the novel SP robotic platform. The patient was discharged from the hospital successfully with no complications during or after the surgery. Pathologic diagnosis showed adenocarcinoma of the gastric mucosa with partial signet-ring cell carcinoma, and no metastasis was found in the 29 cleared lymph nodes. The use of the SHURUI SP robot for total gastrectomy in treating gastric cancer is both technically feasible and safe.

Analysis of short-term ventilation weaning for patients in spontaneous supratentorial intracranial hemorrhage.

Prolonged ventilation is a complication of spontaneous supratentorial hemorrhage patients, but the predictive relationship with successful weaning in this patient cohort is not understood. Here, we evaluate the incidence and factors of ventilation weaning in case of spontaneous supratentorial hemorrhage. We retrospectively studied data from 166 patients in the same hospital from January 2015 to March 2021 and analyzed factors for ventilation weaning. The clinical data recorded included patient age, gender, timing of operation, initial Glasgow Coma Scale (GCS), Intracranial hemorrhage (ICH) score, alcohol drinking, cigarette smoking, medical comorbidity, and the blood data. Predictors of patient outcomes were determined by the Student t test, chi-square test, and logistic regression. We recruited and followed 166 patients who received operation for spontaneous supratentorial hemorrhage with cerebral herniation. The group of successful weaning had 84 patients and the group of weaning failed had 82 patients. The patient's age, type of operation, GCS on admission to the Intensive care unit (ICU), GCS at discharge from the ICU, medical comorbidity was significantly associated with successful weaning, according to Student t test and the chi-square test. According to our findings, patients with stereotaxic surgery, less history of cardiovascular or prior cerebral infarction, GCS >8 before admission to the hospital for craniotomy, and a blood albumin value >3.5 g/dL have a higher chance of being successfully weaned off the ventilator within 14 days.

Well-defined in-textile photolithography towards permeable textile electronics.

Textile-based wearable electronics have attracted intensive research interest due to their excellent flexibility and breathability inherent in the unique three-dimensional porous structures. However, one of the challenges lies in achieving highly conductive patterns with high precision and robustness without sacrificing the wearing comfort. Herein, we developed a universal and robust in-textile photolithography strategy for precise and uniform metal patterning on porous textile architectures. The as-fabricated metal patterns realized a high precision of sub-100 µm with desirable mechanical stability, washability, and permeability. Moreover, such controllable coating permeated inside the textile scaffold contributes to the significant performance enhancement of miniaturized devices and electronics integration through both sides of the textiles. As a proof-of-concept, a fully integrated in-textiles system for multiplexed sweat sensing was demonstrated. The proposed method opens up new possibilities for constructing multifunctional textile-based flexible electronics with reliable performance and wearing comfort.

Research progress of propofol in alleviating cerebral ischemia/reperfusion injury.

Ischemic stroke is a leading cause of adult disability and death worldwide. The primary treatment for cerebral ischemia patients is to restore blood supply to the ischemic region as quickly as possible. However, in most cases, more severe tissue damage occurs, which is known as cerebral ischemia/reperfusion (I/R) injury. The pathological mechanisms of brain I/R injury include mitochondrial dysfunction, oxidative stress, excitotoxicity, calcium overload, neuroinflammation, programmed cell death and others. Propofol (2,6-diisopropylphenol), a short-acting intravenous anesthetic, possesses not only sedative and hypnotic effects but also immunomodulatory and neuroprotective effects. Numerous studies have reported the protective properties of propofol during brain I/R injury. In this review, we summarize the potential protective mechanisms of propofol to provide insights for its better clinical application in alleviating cerebral I/R injury.

Selenium-Mediated Shaping of Citrus Rhizobiome for Promotion in Root Growth and Soil Phosphorus Activation.

Selenium (Se) has been widely reported to affect plant growth, nutrient cycling, and the rhizobiome. However, how Se shapes the rhizobiome and interacts with plants remains largely elusive. Pot and hydroponic experiments were employed to elucidate the regulatory mechanism of Se in the citrus rhizobiome. Compared to the control, soil Se application significantly increased the root biomass (34.7%) and markedly reduced rhizosphere HCl-P, H2O-P, NaHCO3-IP, and residual-P of citrus, which were related to the variation of citrus rhizobiome. Se primarily enriched Proteobacteria and Actinobacteria as well as the phosphorus (P) functional genes phod and pqqc. Further study revealed that Se altered the metabolite profile of root exudate, particularly enhancing the abundance of l-cyclopentylglycine, cycloleucine, l-proline, l-pipecolic acid, and inositol, which played a key role in reshaping the citrus rhizobiome. These metabolites could serve as both nutrient sources and signaling molecules, thus supporting the growth or chemotaxis of the functional microbes. These bacterial taxa have the potential to solubilize P or stimulate plant growth. These findings provide a novel mechanistic understanding of the intriguing interactions between Se, root exudate, and rhizosphere microbiomes, and demonstrate the potential for utilizing Se to regulate rhizobiome function and enhance soil P utilization in citrus cultivation.

A high-permeability method for extracting purple yam saponins based on ultrasonic-assisted natural deep eutectic solvent.

The extraction of active ingredients from traditional Chinese medicine has received considerable attentions. In this study, 16 kinds of natural deep eutectic solvent (NADES) with ultrasonic were selected to extract saponins from purple yam root and the extraction mechanism was investigated. The results showed that chloride/acrylic acid (1:2; n/n) had the highest extraction yield for saponins. The optimal extraction process parameters were 24% water content, 20 mL/g liquid-solid ratio, and ultrasonic extraction for 85 min (81 °C, 600 W). The extraction rate (ER) of purple yam saponins was 0.935%, close to the fitted result of 96.5 mg/g. Molecular dynamics simulations and FT-IR results showed that the NADES may extract the saponin constituents from purple yam through hydrogen bonding. Compared with traditional extraction methods and molecularly imprinted polymer methods, NADES has a higher ER and lower cost (1.53 $/g), which provides a reference for subsequent industrial quantitative production.

Visualizing Macrophage Phenotypes and Polarization in Diseases: From Biomarkers to Molecular Probes.

Macrophage is a kind of immune cell and performs multiple functions including pathogen phagocytosis, antigen presentation and tissue remodeling. To fulfill their functionally distinct roles, macrophages undergo polarization towards a spectrum of phenotypes, particularly the classically activated (M1) and alternatively activated (M2) subtypes. However, the binary M1/M2 phenotype fails to capture the complexity of macrophages subpopulations in vivo. Hence, it is crucial to employ spatiotemporal imaging techniques to visualize macrophage phenotypes and polarization, enabling the monitoring of disease progression and assessment of therapeutic responses to drug candidates. This review begins by discussing the origin, function and diversity of macrophage under physiological and pathological conditions. Subsequently, we summarize the identified macrophage phenotypes and their specific biomarkers. In addition, we present the imaging probes locating the lesions by visualizing macrophages with specific phenotype in vivo. Finally, we discuss the challenges and prospects associated with monitoring immune microenvironment and disease progression through imaging of macrophage phenotypes.

Generalized anxiety disorder in type 2 diabetes mellitus: prevalence and clinical characteristics

Objective: This study investigated the prevalence of generalized anxiety disorder (GAD) in Taiwanese patients with type 2 diabetes mellitus (T2DM). Methods: This retrospective observational study was conducted with a random sample of patients from the entire population of National Health Insurance enrollees during 2000-2010 and used ICD-9-CM diagnostic codes to identify T2DM patients and GAD. The prevalence of GAD was compared between T2DM patients and the general population. Results: Between 2000 and 2010, the prevalence of GAD was significantly greater in the T2DM patients than the general population, while the increase of GAD was higher in the general population (from 0.25 to 0.63%) than among T2DM patients (from 0.81 to 1.03%). In T2DM patients, GAD was associated with female gender, a Charlson Comorbidity Index ≥ 1, diabetes mellitus duration > 9 years, and the following comorbidities: congestive heart failure, peripheral vascular disease, and depressive disorder. The prevalence of GAD among T2DM patients was negatively associated with rapid-acting insulin injection therapy and with the use of metformin and sulfonylureas. Conclusion: Since the prevalence of GAD was greater among T2DM patients than the general population, public health initiatives are needed to prevent and treat GAD in T2DM patients, specifically those with the above mentioned risk factors.