Delta opioid peptide [D-ala2, D-leu5]-Enkephalin's ability to enhance mitophagy via TRPV4 to relieve ischemia/reperfusion injury in brain microvascular endothelial cells.

BACKGROUND: Local brain tissue can suffer from ischaemia/reperfusion (I/R) injury, which lead to vascular endothelial damage. The peptide δ opioid receptor (δOR) agonist [D-ala2, D-leu5]-Enkephalin (DADLE) can reduce apoptosis caused by acute I/R injury in brain microvascular endothelial cells (BMECs). OBJECTIVE: This study aims to explore the mechanism by which DADLE enhances the level of mitophagy in BMECs by upregulating the expression of transient receptor potential vanilloid subtype 4 (TRPV4). METHODS: BMECs were extracted and made to undergo oxygen-glucose deprivation/reoxygenation (OGD/R) accompanied by DADLE. RNA-seq analysis revealed that DADLE induced increased TRPV4 expression. The CCK-8 method was used to assess the cellular viability; quantitative PCR (qPCR) was used to determine the mRNA expression of Drp1; western blot was used to determine the expression of TRPV4 and autophagy-related proteins; and calcium imaging was used to detect the calcium influx. Autophagosomes in in the cells' mitochondria were observed by using transmission electron microscopy. ELISA was used to measure ATP content, and a JC-1 fluorescent probe was used to detect mitochondrial membrane potential. RESULTS: When compared with the OGD/R group, OGD/R+DADLE group showed significantly enhanced cellular viability; increased expression of TRPV4, Beclin-1, LC3-II/I, PINK1 and Parkin; decreased p62 expression; a marked rise in calcium influx; further increases in mitophagy, an increase in ATP synthesis and an elevation of mitochondrial membrane potential. These protective effects of DADLE can be blocked by a TRPV4 inhibitor HC067047 or RNAi of TRPV4. CONCLUSION: DADLE can promote mitophagy in BMECs through TRPV4, improving mitochondrial function and relieving I/R injury.

Fundamental, mechanism and development of hydration lubrication: From bio-inspiration to artificial manufacturing.

Friction and lubrication are ubiquitous in all kinds of movements and play a vital role in the smooth operation of production machinery. Water is indispensable both in the lubrication systems of natural organisms and in hydration lubrication systems. There exists a high degree of similarity between these systems, which has driven the development of hydration lubrication from biomimetic to artificial manufacturing. In particular, significant advancements have been made in the understanding of the mechanisms of hydration lubrication over the past 30 years. This enhanced understanding has further stimulated the exploration of biomimetic inspiration from natural hydration lubrication systems, to develop novel artificial hydration lubrication systems that are cost-effective, easily transportable, and possess excellent capability. This review summarizes the recent experimental and theoretical advances in the understanding of hydration-lubrication processes. The entire paper is divided into three parts. Firstly, surface interactions relevant to hydration lubrication are discussed, encompassing topics such as hydrogen bonding, hydration layer, electric double layer force, hydration force, and Stribeck curve. The second part begins with an introduction to articular cartilage in biomaterial lubrication, discussing its compositional structure and lubrication mechanisms. Subsequently, three major categories of bio-inspired artificial manufacturing lubricating material systems are presented, including hydrogels, polymer brushes (e.g., neutral, positive, negative and zwitterionic brushes), hydration lubricant additives (e.g., nano-particles, polymers, ionic liquids), and their related lubrication mechanism is also described. Finally, the challenges and perspectives for hydration lubrication research and materials development are presented.

Histopathological staging and differential diagnosis of marginal zone lymphoma of gastric mucosa-associated lymphoid tissue.

The purpose of this study was to explore the histopathological staging and differential diagnosis of marginal zone lymphoma in gastric mucosa-associated lymphoid tissue (MALT lymphoma). We performed detailed histomorphology and immunohistochemistry investigations as well as genetic testing on endoscopic biopsy and endoscopic mucosal resection specimens from 18 patients with gastric MALT lymphoma. We found that gastric MALT lymphoma typically begins as a small, isolated area outside the lymphoid follicular mantle zone or proliferates in a multifocal, patchy manner, gradually spreads to the interfollicular zone, forming diffuse proliferation, invades the gastric mucosal glands, and infiltrates or proliferates into the center of peripheral reactive lymphoid follicles. Abnormally proliferating lymphocytes invade the surrounding lymphoid follicles, resulting in damage, atrophy, and disappearance of their normal follicles as well as of the gastric mucosa glands, forming diffuse proliferation. Redifferentiation and proliferation lead to the transformation of lymphocytes; that is, MALT transitions into highly invasive lymphoma. Based on our findings in this study, we propose the following five stages in the process of development and progression of gastric MALT lymphoma: the stage of cell proliferation outside the lymphoid follicular mantle zone; the stage of heterogeneous proliferative lymphoepithelial lesion; the stage of reactive lymphoid follicular implantation; the stage of lymphoid follicular clonal proliferation; and the stage of MALT transforming into highly invasive lymphoma. We examined the differential diagnosis of histopathological features at each stage. The clinicopathological staging of gastric MALT lymphoma can help clinicians provide accurate treatment and track malignant cell transformation, thus playing a significant role in controlling its development and progression.

Tibial cortex transverse transport regulates Orai1/STIM1-mediated NO release and improve the migration and proliferation of vessels via increasing osteopontin expression.

Background: Diabetic foot is a major complication of diabetes. The bone transverse transport method could be applied in clinics for treatment, which could improve the metabolism of the tissues via lasting distraction forces. However, the process' specific regulating mechanism is still unknown. Methods: Based on the notion that the healing of bones involves the recruitment of calcium ions, in this study, we established the model of tibial cortex transverse transport (TTT) on rats and then used tissue immunologic detection, such as the double fluorescent staining to explore the expression of the calcium channels' calcium release-activated calcium modulator 1 (Orai1)/stromal interaction molecule 1 (STIM1), which belong to the store-operated calcium entry (SOCE) signaling pathways on the tissues around the bone transport area. By using the laser capture microdissection (LCM) tool, we acquired samples of tissues around the bone and endeavored to identify pivotal protein molecules. Subsequently, we validated the functions of key protein molecules through in vitro and in vivo experiments. Results: After protein profile analysis, we found the differentially expressed key protein osteopontin (OPN). The in vitro experiments verified that, being stimulated by OPN, the migration, proliferation, and angiogenesis of human umbilical vein endothelial cells (HUVEC) were observed to be enhanced. The activation of Orai1/STIM1 might increase the activity of endothelial nitric oxide synthase (eNOS) and its effect on releasing nitric oxide (NO). Subsequently, the migration and proliferation of the HUVECs are improved, which ultimately accelerates wound healing. These signaling pathway was also observed in the OPN-stimulated healing process of the skin wound surface of diabetic mice. Conclusion: This study identifies the molecular biological mechanism of OPN-benefited the migration and proliferation of the HUVECs and provides ideas for searching for new therapeutic targets for drugs that repair diabetes-induced wounds to replace invasive treatment methods. The translational potential of this article: The OPN is highly expressed in the tissues surrounding the TTT bone transfer area, which may possibly stimulate the activation of eNOS to increase NO release through the SOCE pathway mediated by Orai1/STIM1. This mechanism may play a significant role in the angiogenesis of diabetic foot's wounds promoted by TTT, providing new therapeutic strategies for the non-surgical treatment for this disease.

Effects of Heat Treatment on Microstructures and Mechanical Properties of a Low-Alloy Cylinder Liner.

Cylinder liners, considered a crucial component of internal combustion (IC) engines, often require excellent mechanical properties to ensure optimal engine performance under elevated temperatures, pressures, and varying loads. In this work, a new low-alloy cylinder liner, incorporating a low content of molybdenum, copper, and chromium into gray cast iron, was fabricated using a centrifugal casting process. Subsequently, the heat treatment processes were designed to achieve bainite microstructures in the cylinder liner through rapid air cooling, isothermal transformation, and tempering. The effects of different air-cooling rates and tempering temperatures on the microstructure evolution and mechanical properties of cylinder liner were investigated. The results revealed that during the supercooled austenite transformation process, rapid air cooling at a rate of 14.5-23.3 °C/s can effectively bypass the formation of pearlitic structures and directly induce the formation of bainite structures. Once the temperature exceeded 480-520 °C, hardness and tensile strength increased with the temperature increase owing to the enhancement of the lower bainite content, the reduction of residual austenite, and the precipitation of the fine hard carbides in the matrix. With temperatures above 520-550 °C, the carbide and lower bainite organization coarsened, thereby reducing the hardness and tensile strength of the material. Therefore, the optimal heat treatment parameters were rapid cooling at 14.5-23.3 °C/s rate to obtain bainite, and tempering of 480-520 °C for finer and more uniform bainite. In addition, the results of the characterization of the mechanical properties of the cylinder liner after heat treatment showed that the hardness, tensile strength, and wear resistance were improved with the refinement of the bainite.

Multi-omics analysis of functional substances and expression verification in cashmere fineness.

BACKGROUND: Numerous factors influence the growth and development of cashmere. Existing research on cashmere has predominantly emphasized a single omics level. Integrating multi-omics analyses can offer a more comprehensive understanding by encompassing the entire spectrum. This study more accurately and comprehensively identified the key factors influencing cashmere fineness using multi-omics analysis. METHODS: This study used skin tissues of coarse cashmere type (CT_LCG) and fine cashmere type Liaoning cashmere goats (FT_LCG) for the analysis. This study employed an integrated approach involving transcriptomics, translatomics, proteomics, and metabolomics to identify substances associated with cashmere fineness. The findings were validated using parallel reaction monitoring (PRM) and multiple reaction monitoring (MRM) techniques. RESULTS: The GO functional enrichment analysis identified three common terms: multicellular organismal process, immune system process, and extracellular region. Furthermore, the KEGG enrichment analysis uncovered the involvement of the arachidonic acid metabolic pathway. Protein expression trends were verified using PRM technology. The expression trends of KRT79, as confirmed by PRM, were consistent with those observed in TMT proteomics and exhibited a positive regulatory effect on cashmere fineness. Metabolite expression trends were confirmed using MRM technology. The expression trends of 9 out of 15 validated metabolites were in agreement with those identified in the non-targeted metabolomics analysis. CONCLUSIONS: This study employed multi-omics analysis to identify key regulators of cashmere fineness, including PLA2G12A, KRT79, and prostaglandin B2. The findings of this study offer valuable data and establish a theoretical foundation for conducting comprehensive investigations into the molecular regulatory mechanisms and functional aspects of cashmere fineness.

Research on Wind Turbine Fault Detection Based on the Fusion of ASL-CatBoost and TtRSA.

The internal structure of wind turbines is intricate and precise, although the challenging working conditions often give rise to various operational faults. This study aims to address the limitations of traditional machine learning algorithms in wind turbine fault detection and the imbalance of positive and negative samples in the fault detection dataset. To achieve the real-time detection of wind turbine group faults and to capture wind turbine fault state information, an enhanced ASL-CatBoost algorithm is proposed. Additionally, a crawling animal search algorithm that incorporates the Tent chaotic mapping and t-distribution mutation strategy is introduced to assess the sensitivity of the ASL-CatBoost algorithm toward hyperparameters and the difficulty of manual hyperparameter setting. The effectiveness of the proposed hyperparameter optimization strategy, termed the TtRSA algorithm, is demonstrated through a comparison of traditional intelligent optimization algorithms using 11 benchmark test functions. When applied to the hyperparameter optimization of the ASL-CatBoost algorithm, the TtRSA-ASL-CatBoost algorithm exhibits notable enhancements in accuracy, recall, and other performance measures compared with the ASL-CatBoost algorithm and other ensemble learning algorithms. The experimental results affirm that the proposed algorithm model improvement strategy effectively enhances the wind turbine fault detection classification recognition rate.

Nutritional and volatile profiles of pulp and flavedo from four local pummelo cultivars grown in Fujian province of China.

The nutritional and volatile profiles of pulp and flavedo samples from four distinct local pummelo landraces ("Siji," "Pingshan," "Wendan," and "Guanxi") cultivated in Fujian province of China were investigated. "Guanxi" pummelo exhibited relatively high contents of vitamin C (42.01 mg/100 mL) and phenols (360.61 mg/L) and displayed a robust antioxidant capacity (41.15 mg/100 mL). Conversely, the red pulp from "Pingshan" demonstrated relatively high values of carotenoids (55.96 µg/g) and flavonoids (79.79 mg/L). Considerable differences were observed in volatile compositions between the two fruit tissues and among the four genotypes. A total of 166 and 255 volatile compounds were detected in the pulp and flavedo samples, respectively. Notably, limonene and ß-myrcene were identified as the principal volatile compounds in flavedo, whereas hexanal was highly abundant in the pulp of "Siji," "Pingshan," and "Guanxi." "Wendan" displayed distinct separation from the other three pummelo cultivars in principal component analysis based on the pulp volatile compositions. This distinction was attributed to the higher number and content of volatile compounds in "Wendan" pulp, particularly the remarkable enrichment of ß-myrcene. The newly characterized pummelo landraces and genotype/tissue-dependent variations in volatiles provide essential information for the genetic improvement of pummelo aroma, as well as for fruit processing and utilization.

An ultralight and flexible nanofibrillated cellulose/chitosan aerogel for efficient chromium removal: Adsorption-reduction process and mechanism.

Heavy metals in water are critical global environmental problems. In particular, the anionic heavy metal chromium (Cr) has carcinogenic and genotoxic risks on human health. To this end, an ultralight and flexible nanofibrillated cellulose (NFC)/chitosan (CS) aerogel was developed only by freeze-drying combined with physical thermal cross-linking for efficient one step co-removal of Cr(VI) and Cr(III). The maximum adsorption capacity of Cr(VI) and total Cr calculated according to the Langmuir model was 197.33 and 134.12 mg/g, respectively. Even in the presence of competing soluble organics, anions and oil contaminants, the resulting NFC/CS-5 aerogels showed excellent selectivity. The aerogel exhibited outstanding mechanical integrity, remaining intact after 17 compressions in air and underwater. Meanwhile, after 5 adsorption-desorption cycles, the aerogel was easy to regenerate and maintained a high regeneration efficiency of 80.25%. Importantly, self-assembled NFC/CS-5 aerogel filter connected with the peristaltic pump could purify 752 mL of industrial wastewater with Cr(VI) pre-concentration capacity of 49.71 mg/g. XPS and FT-IR verified that electrostatic interactions, reduction and complexation acted as the main driving forces for the adsorption process. Moreover, such aerogel possessed broad application prospects for alleviating heavy metal pollution in agriculture.

Functionally-designed floatable amino-modified ZnLa layered double hydroxides/cellulose acetate beads for tetracycline removal: Performance and mechanism.

The over-reliance on tetracycline antibiotics (TC) in the animal husbandry and medical field has seriously affected the safety of the ecological environment. Therefore, how to effectively treat tetracycline wastewater has always been a long-term global challenge. Here, we developed a novel polyethyleneimine (PEI)/Zn-La layered double hydroxides (LDH)/cellulose acetate (CA) beads with cellular interconnected channels to strengthen the TC removal. The results of the exploration on its adsorption properties illustrated that the adsorption process exhibited a favorable correlation with the Langmuir model and the pseudo-second-order kinetic model, namely monolayer chemisorption. Among the many candidates, the maximum adsorption capacity of TC by 10 %PEI-0.8LDH/CA beads was 316.76 mg/g. Apart from that, the effects of pH, interfering species, actual water matrix and recycling on the adsorption of TC by PEI-LDH/CA beads were also analyzed to verify their superior removal capability. The potential for industrial-scale applications was expanded through fixed-bed column experiments. The proven adsorption mechanisms mainly included electrostatic interaction, complexation, hydrogen bonding, n-π EDA effect and cation-π interaction. The self-floating high-performance PEI-LDH/CA beads exploited in this work provided fundamental support for the practical application of antibiotic-based wastewater treatment.

A critical review on correlating active sites, oxidative species and degradation routes with persulfate-based antibiotics oxidation.

At present, numerous heterogeneous catalysts have been synthesized to activate persulfate (PS) and produce various reactive species for antibiotic degradation from water. However, the systematic summary of the correlation among catalyst active sites, PS activation pathway and pollutant degradation has not been reported. This review summarized the effect of metal-based, carbon-based and metal-carbon composite catalysts on the degradation of antibiotics by activating PS. Metal and non-metal sites are conducive to inducing different oxidation pathways (SO4•-, •OH radical oxidation and 1O2 oxidation, mediated electron transfer, surface-bound reactive complexes and high-valent metal oxidation). SO4•- and •OH are easy to attack CH, S-N, CN bonds, CC double bonds and amino groups in antibiotics. 1O2 is more selective to the structure of the aniline ring and amino group, and also to attacking CS, CN and CH bonds. Surface-bound active species can cleave CC, SN, CS and CN bonds. Other non-radical pathways may also induce different antibiotic degradation routes due to differences in oxidation potential and electronic properties. This critical review clarified the functions of active sites in producing different reactive species for selective oxidation of antibiotics via featured pathways. The outcomes will provide valuable guidance of oriented-regulation of active sites in heterogeneous catalysts to produce on-demand reactive species toward high-efficiency removing antibiotics from water.

Does Preoperative Hookwire Localization Influence Postoperative Acute and Chronic Pain After Video-Assisted Thoracoscopic Surgery: A Prospective Cohort Study.

Purpose: This study aimed to investigate whether preoperative computerized tomography-guided hookwire localization-associated pain could affect acute and chronic postsurgical pain (CPSP) in patients undergoing video-assisted thoracoscopic surgery (VATS). Methods: We enrolled 161 adult patients who underwent elective VATS; sixty-nine patients experienced hookwire localization (Group A) and 69 did not (Group B). Group A was further subdivided into the multiple localization group (n=35, Group Amultiple) and the single localization group (n=34, Group Asingle) according to the number of hookwires. The numerical rating scale (NRS) was used preoperatively, during recovery at the post-anesthesia care unit (PACU), and the first two days, 3 and 6 months postoperatively. Furthermore, multivariate regression analysis was used to identify the risk factors associated with CPSP. The postoperative adverse events, length of hospital stay, and satisfaction in pain management were also recorded. Results: The incidence and severity of acute postoperative pain were similar between Group A and Group B (p > 0.05). The incidence (56.5% vs 30.4%, p = 0.002) and the NRS scores (2.0 [2.0-3.0] vs 1.0 [1.0-2.0], p = 0.011) for CPSP were significantly higher in Group A than in Group B at 3 months postoperatively. On subgroup analysis, compared with Group Asingle, the intensity of CPSP (2.0 [2.0-3.0] vs 2.0 [1.0-2.0], p = 0.005) in Group Amultiple was slightly higher at 3 months postoperatively. Conversely, the CPSP incidence (60.0% vs 29.4%, p = 0.011) was significantly higher at 6 months postoperatively in Group Amultiple. The multivariate regression analysis further validated hookwire localization as a risk factor for CPSP (odds ratio: 6.199, 95% confidence interval 2.049-18.749, p = 0.001). Patient satisfaction relating to pain management at 3 months postoperatively was lower in Group A (p = 0.034). Conclusion: The preoperative pain stress of hookwire localization increased the incidence and intensity of CPSP rather than acute pain at 3 months postoperatively, especially in patients with multiple hookwires.

Application of MXene-based materials in Fenton-like systems for organic wastewater treatment: A review.

Recently, Fenton-like systems have been widely explored and applied for the removal of organic matter from wastewater. Two-dimensional (2D) MXene-based materials exhibit excellent adsorption and catalysis capacity for organic pollutants removal, which has been reported widely. However, there is no summary on the application of MXene-based materials in Fenton-like systems for organic matter removal. In this review, four types of MXene-based materials were introduced, including 2D MXene, MXene/Metal complex, MXene/Metal oxide complex, and MXene/3D carbon material complex. In addition, the Fenton-like system usually consists of adsorption and degradation processes. The oxidation process might contain hydrogen peroxide (H2O2) or persulfate (PS) oxidants. This review summarizes the performance and mechanisms of organic pollutants adsorption and oxidants activation by MXene-based materials systematically. Finally, the existing problems and future research directions of MXene-based materials are proposed in Fenton-like wastewater treatment systems.

In situ identification of environmental microorganisms with Raman spectroscopy.

Microorganisms in natural environments are crucial in maintaining the material and energy cycle and the ecological balance of the environment. However, it is challenging to delineate environmental microbes' actual metabolic pathways and intraspecific heterogeneity because most microorganisms cannot be cultivated. Raman spectroscopy is a culture-independent technique that can collect molecular vibration profiles from cells. It can reveal the physiological and biochemical information at the single-cell level rapidly and non-destructively in situ. The first part of this review introduces the principles, advantages, progress, and analytical methods of Raman spectroscopy applied in environmental microbiology. The second part summarizes the applications of Raman spectroscopy combined with stable isotope probing (SIP), fluorescence in situ hybridization (FISH), Raman-activated cell sorting and genomic sequencing, and machine learning in microbiological studies. Finally, this review discusses expectations of Raman spectroscopy and future advances to be made in identifying microorganisms, especially for uncultured microorganisms.

Self-supported trimetallic NiZnLa nanosheets on hierarchical porous graphene oxide-polymer composite fibers for enhanced phosphate removal from water.

Phosphate-induced water eutrophication has attracted global attention. Fabricating adsorbents with both high phosphate adsorption affinity and accessible separation property is challenging. Herein, PG@NZL, a hierarchical nanocomposite fibrous membrane, was fabricated via in-situ growth of La-doped NiZn-LDH (NiZnLa0.1) over electrospun graphene oxide-polymer composite fibers (PG). The porous surface of the PG fibers provided abundant anchor sites for the vertical self-supported growth of NiZnLa0.1 nanosheets, contributing to a high surface area. The La-doped NiZnLa0.1 trimetallic LDH achieved a much higher adsorption capacity than NiZn-LDH. The negative adsorption energy (-1.45 eV), calculated with DFT, confirmed its spontaneous adsorption potential for phosphate. Interestingly, the PG fibers contributed to oxygen vacancies and the metal center electronic structure evolution of NiZnLa0.1, thus strengthening the coordination with phosphate. Mechanistic analysis revealed that the high adsorption capacity of PG@NZL is attributed to its superior anion exchange property, oxygen vacancies, and inner-sphere complexation. Therefore, the flexible and easily separated PG@NZL nanocomposite fibrous membrane is a promising adsorbent for effectively treating phosphate-bearing wastewater.

Bowknot-like Zr/La bimetallic organic frameworks for enhanced arsenate and phosphate removal: Combined experimental and DFT studies.

Hazardous oxyanions in water bodies are potentially toxic to aquatic life, and the coexistence of multiple anions aggravates the toxicity. Herein, bowknot-like Zr/La bimetallic organic frameworks (Zr/La-BTC) were developed with superior hazardous oxyanion adsorption capacities, i.e., approximately 102 mg/g for arsenate and 159 mg/g for phosphate, respectively. The molar ratio of Zr to La in Zr/La-BTC plays a significant role in the structure and the adsorption efficiencies. Notably, the experiment-derived adsorption capacities of various Zr/La-BTC samples were consistent with their adsorption energies calculated by density-function theory (DFT). Further mechanism analysis revealed that coordination of Zr/La atoms with the target anion groups occurred during adsorption. The positive shift of binding energies in La 3d and Zr 3d XPS spectra and Bader charge analysis unveiled that back-donation interactions dominated the adsorption process. The reliable adsorption selectivity and reusability of 0.1Zr/La-BTC were verified with anion competition experiments and four adsorption-desorption cycles. Overall, this study provides significant insight into the design of high-performance bimetallic organic frameworks for the enhanced removal of hazardous oxyanions from water.

Fruit quality and volatile constituents of a new very early-ripening pummelo (Citrus maxima) cultivar 'Liuyuezao'.

'Liuyuezao' (LYZ) pummelo (Citrus maxima) originated from a spontaneous bud sport on a 'Guanxi' (GXB) pummelo tree and was released as a new very early-season cultivar. The objective of this study was to present the sensory and nutritional profiles of LYZ fruits, and compare it with other major commercialized pummelo cultivars including GXB, 'Sanhong' (SH) and 'Hongrou' (HR). LYZ had higher contents of organic acids (12.01 mg/g), phenols (669.01 mg/L), vitamin C (75.73 mg/100 mL) and stronger antioxidant capacity (77.65 mg/100 mL) but lower levels of soluble sugars (62.85 mg/g), carotenoids (0.25 mg/L) and flavonoids (46.3 mg/L) when compared to the other pummelos. Moreover, a smaller number (49) and much less content (7.63) of fruit volatiles were detected in LYZ than them in GXB, SH and HR. The relatively high levels of fructose (20.6 mg/g) and organic acids and low levels of volatile compounds in LYZ mainly contributed to its sweet and mildly sour taste and moderate aroma of pummelo note. LYZ is presented as an alternative pummelo cultivar with the potential for commercialization.

Delta Opioid Peptide Targets Brain Microvascular Endothelial Cells Reducing Apoptosis to Relieve Hypoxia-Ischemic/Reperfusion Injury.

Stroke is one of the leading causes of death. (D-ala2, D-leu5) enkephalin (DADLE) is a synthetic peptide and highly selective delta opioid receptor (δOR) agonist that has exhibited protective properties in ischemia. However, the specific target and mechanism are still unclear. The present study explores the expression of δOR on brain microvascular endothelial cells (BMECs) and whether DADLE could relieve I/R-induced injury by reducing apoptosis. A lateral ventricular injection of DADLE for pretreatment, the neurofunctional behavior score, and TTC staining, were used to evaluate the protective effect of DADLE. Immunofluorescence technology was used to label different types of cells with apoptosis-positive signals to test co-localization status. Primary cultured BMECs were separated and treated with DADLE, accompanied by OGD/R. The CCK-8 test was conducted to evaluate cell viability and TdT-mediated dUTP Nick-end Labelling (TUNEL) staining to test apoptosis levels. The levels of apoptosis-related proteins were analyzed by Western blotting. The co-localization results showed that BMECs, but not astrocytes, microglia, or neurons, presented mostly TUNEL-positive signals, especially in the Dentate gyrus (DG) area of the hippocampus. Either activation of δORs on rats' brains or primary BMECs mainly reduce cellular apoptosis and relieve the injury. Interference with the expression δOR could block this effect. DADLE also significantly increased levels of Bcl-2 and reduced levels of Bax. δOR's expressions can be detected on the BMECs, but not on the HEK293 cells, by Western blotting and IFC. Therefore, DADLE exerts a cytoprotective effect, primarily under hypoxia-ischemic injury/reperfusion conditions, by targeting BMECs to inhibit apoptosis.

Classification and Identification of Archaea Using Single-Cell Raman Ejection and Artificial Intelligence: Implications for Investigating Uncultivated Microorganisms.

Archaea can produce special cellular components such as polyhydroxyalkanoates, carotenoids, rhodopsin, and ether lipids, which have valuable applications in medicine and green energy production. Most of the archaeal species are uncultivated, posing challenges to investigating their biomarker components and biochemical properties. In this study, we applied Raman spectroscopy to examine the biological characteristics of nine archaeal isolates, including halophilic archaea (Haloferax larsenii, Haloarcula argentinensis, Haloferax mediterranei, Halomicrobium mukohataei, Halomicrobium salinus, Halorussus sp., Natrinema gari), thermophilic archaea (Sulfolobus acidocaldarius), and marine group I (MGI) archaea (Nitrosopumilus maritimus). Linear discriminant analysis of the Raman spectra allowed visualization of significant separations among the nine archaeal isolates. Machine-learning classification models based on support vector machine achieved accuracies of 88-100% when classifying the nine archaeal species. The predicted results were validated by DNA sequencing analysis of cells isolated from the mixture by Raman-activated cell sorting. Raman spectra of uncultured archaea (MGII) were also obtained based on Raman spectroscopy and fluorescence in situ hybridization. The results combining multiple Raman-based techniques indicated that MGII may have the ability to produce lipids distinct from other archaeal species. Our study provides a valuable approach for investigating and classifying archaea, especially uncultured species, at the single-cell level.