Engineering of Halide Methyltransferase BxHMT through Dynamic Cross-Correlation Network Analysis.

Halide methyltransferases (HMTs) provide an effective way to regenerate S-adenosyl methionine (SAM) from S-adenosyl homocysteine and reactive electrophiles, such as methyl iodide (MeI) and methyl toluene sulfonate (MeOTs). As compared with MeI, the cost-effective unnatural substrate MeOTs can be accessed directly from cheap and abundant alcohols, but shows only limited reactivity in SAM production. In this study, we developed a dynamic cross-correlation network analysis (DCCNA) strategy for quickly identifying hot spots influencing the catalytic efficiency of the enzyme, and applied it to the evolution of HMT from Paraburkholderia xenovorans. Finally, the optimal mutant, M4 (V55T/C125S/L127T/L129P), exhibited remarkable improvement, with a specific activity of 4.08 U/mg towards MeOTs, representing an 82-fold increase as compared to the wild-type (WT) enzyme. Notably, M4 also demonstrated a positive impact on the catalytic ability with other methyl donors. The structural mechanism behind the enhanced enzyme activity was uncovered by molecular dynamics simulations. Our work not only contributes a promising biocatalyst for the regeneration of SAM, but also offers a strategy for efficient enzyme engineering.

Polarity-Ultrasensitive and Lipophilicity-Enhanced Structurally Modified Hemicyanine for Two-Color Staining to Reveal Cell Apoptosis during Chemotherapy.

Programmed cell death (PCD) is a precisely controlled physiological process to sustain tissue homeostasis. Even though the PCD pathways have been explicitly subdivided, the individual cell death process seems to synergistically operate to eliminate cells rather than separately execute signal transduction. Apoptosis is the dominant intracellular PCD subtype, which is intimately regulated and controlled by mitochondria, thus tracing mitochondrial actions could reveal the dynamic changes of apoptosis, which may provide important tools for screening preclinical therapeutic agents. Herein, we exploited an innovative fluorophore Cy496 based on the light-initiated cleavage reaction. Cy496 bears the typical D-π-A structure and serves as a versatile building block for chemosensor construction through flexible side chains. By regulating lipophilicity and basicity through bis-site substitution, we synthesized a series of fluorescence probes and screened a novel mitochondria-targeted ratiometric probe Cy1321, which can real-time evaluate the dynamic changes of mitochondrial micropolarity mediated by bis-cholesterol anchoring. Cy1321 has realized two-color quantification and real-time visualization of polarity fluctuations on chemotherapy agent (cisplatin)-induced apoptosis through flow cytometry and confocal imaging and also achieved the purpose of detecting mitochondria-related apoptosis at the level of tissues. It is envisioned that Cy1321 has sufficient capability as a promising and facile tool for the evaluation of apoptosis and contributing to therapeutic drug screening.

Analysis of the Development Status and Prospect of China's Agricultural Sensor Market under Smart Agriculture.

Agricultural sensors are essential technologies for smart agriculture, which can transform non-electrical physical quantities such as environmental factors. The ecological elements inside and outside of plants and animals are converted into electrical signals for control system recognition, providing a basis for decision-making in smart agriculture. With the rapid development of smart agriculture in China, agricultural sensors have ushered in opportunities and challenges. Based on a literature review and data statistics, this paper analyzes the market prospects and market scale of agricultural sensors in China from four perspectives: field farming, facility farming, livestock and poultry farming and aquaculture. The study further predicts the demand for agricultural sensors in 2025 and 2035. The results reveal that China's sensor market has a good development prospect. However, the paper garnered the key challenges of China's agricultural sensor industry, including a weak technical foundation, poor enterprise research capacity, high importation of sensors and a lack of financial support. Given this, the agricultural sensor market should be comprehensively distributed in terms of policy, funding, expertise and innovative technology. In addition, this paper highlighted integrating the future development direction of China's agricultural sensor technology with new technologies and China's agricultural development needs.

Homocysteine-specific fluorescence detection and quantification for evaluating S-adenosylhomocysteine hydrolase activity.

Studies have shown that homocysteine (Hcy) levels are closely related to cardiovascular and cerebrovascular diseases. In this work, we have developed and synthesized three copper complexes, F542-Cu2+, F508-Cu2+, and F465-Cu2+ for Hcy detection. The different binding constants (Ks) of the copper complexes endow them with dramatic reactivity toward biothiols. The pyridine-containing tetraazacycle was employed in the construction of F542-Cu2+, which renders the medium Ks value for the copper complex compared with cyclen and TACN and effectively prevented the disintegration of the complexes. Pyridine-containing tetraazacycle provided the basis and possibility for the hypothesis for the reduction of Cu2+ by biothiols to shape into a stable six-membered ring structure. The obtained results verified that F542-Cu2+ could be utilized to specifically probe Hcy in a switched-on fluorescence mode. F542-Cu2+ exhibited excellent environmental stability, superior sensitivity, and outstanding selectivity toward Hcy under physiological conditions. The mechanism of Hcy specificity was confirmed to be related to the generation of Hcy-induced six-membered ring by fluorescence imaging, time-dependent fluorescence spectra, ESI-MS, and electron paramagnetic resonance (EPR) analyses. Furthermore, we exploited the application of F542-Cu2+ and developed a strategy for evaluating the activity of S-adenosylhomocysteine hydrolase (AHCY) in vitro by fluorescence analysis. More importantly, real-time in vivo evaluation of the enzymatic activity of AHCY was realized and assisted by our probe, providing the possibility of opening up a new avenue for enzymatic reaction assessment.

The increased motion of lumbar induces ligamentum flavum hypertrophy in a rat model.

BACKGROUND: The purpose of this study was to establish a novel rat model for ligamentum flavum (LF) hypertrophy using increased motion of lumbar and to elucidate the etiology of (LFH). METHODS: A total number of 30 male rats were used. The increased motion of lumbar was induced by surgical resection of L5/6 posterior elements (n = 15). The other rats underwent a sham operation (n = 15). After 8 weeks, all rats were taken lateral plain X-rays. The LF from L5/6 in both groups were harvested to investigate histological, immunohistological, and real-time PCR analysis. RESULTS: According to radiological results, the disc height ratio, flexion ratio, and extension ratio were larger in the rats in the experimental group than that of in the sham group. The HE staining showed that the LF thickness in the experimental group significantly increased in comparison to the sham group. The Masson trichrome staining showed that the ratio of elastic fibers to collagen fibers in experimental group was lower than that in the sham group. The protein and gene expression of TGF-ß1, TNF-α, IL-1ß, and Col 1 were significantly higher in the experimental group than that in the sham group. CONCLUSION: A relatively safe, simple, and rapid rat model of LFH using increased motion of lumbar was established. The increased motion of lumbar could lead to high expression of inflammatory and fibrotic factors in LF, causing the accumulation of collagen fibers and decreasing of elastic fibers.

Antimicrobial Mechanism of pBD2 against Staphylococcus aureus.

Antimicrobial peptides (AMPs) show high antibacterial activity against pathogens, which makes them potential new therapeutics to prevent and cure diseases. Porcine beta defensin 2 (pBD2) is a newly discovered AMP and has shown antibacterial activity against different bacterial species including multi-resistant bacteria. In this study, the functional mechanism of pBD2 antibacterial activity against Staphylococcus aureus was investigated. After S. aureus cells were incubated with different concentrations of pBD2, the morphological changes in S. aureus and locations of pBD2 were detected by electron microscopy. The differentially expressed genes (DEGs) were also analyzed. The results showed that the bacterial membranes were broken, bulging, and perforated after treatment with pBD2; pBD2 was mainly located on the membranes, and some entered the cytoplasm. Furthermore, 31 DEGs were detected and confirmed by quantitative real-time PCR (qRT-PCR). The known functional DEGs were associated with transmembrane transport, transport of inheritable information, and other metabolic processes. Our data suggest that pBD2 might have multiple modes of action, and the main mechanism by which pBD2 kills S. aureus is the destruction of the membrane and interaction with DNA. The results imply that pBD2 is an effective bactericide for S. aureus, and deserves further study as a new therapeutic substance against S. aureus.

Study on the mechanism of action of Wu Mei Pill in inhibiting rheumatoid arthritis through TLR4-NF-κB pathway.

BACKGROUND: Wu Mei Pills (WMP) is a traditional Chinese medication that exhibits considerable anti-inflammatory effects. While WMP has been documented for its efficacy in treating RA, its mechanism of action on the condition remains unestablished. METHODS: The chemical composition of WMP was analyzed through UPLC-MS. Next, the enzyme-linked immunosorbent assay, cell scratch, Transwell, and Western blotting techniques were used to investigate its intrinsic mechanism. Lastly, the effect of WMP in inhibiting RA was explored by applying it to CIA rats. RESULT: UPLC-MS analysis detected 181 compounds in WMP. RA-FLS migration and invasion mechanisms were significantly hindered by serum containing WMP (2%, 8%). Moreover, WMP (0.5 g/kg, 2 g/kg) restricted arthritis and immune organ indices in CIA rats with type II collagen-induced rheumatoid arthritis by blocking TLR4-NF-κB inflammatory pathway activation. CONCLUSIONS: WMP is valuable in mitigating the course of RA through inhibiting the classical TLR4-NF-κB inflammatory pathway and reducing the secretion of inflammatory factors in the serum of RA-FLS and CIA rats. Moreover, it regulates the dynamic balance of MMP-2/TIMP-2, MMP-9/TIMP-1, modulates the mechanism of RA-FLS invasion, and safeguards articular cartilage tissues in RA.

Quantification of Allergic Crustacean Tropomyosin Using Shared Signature Peptides in Processed Foods with a Mass Spectrometry-Based Proteomic Strategy.

Crustacean shellfish are major allergens in East Asia. In the present study, a major allergic protein in crustaceans, tropomyosin, was detected accurately using multiple reaction monitoring mode-based mass spectrometry, with shared signature peptides identified through proteomic analysis. The peptides were deliberately screened through thermal stability and enzymatic digestion efficiency to improve the suitability and accuracy of the developed method. Finally, the proposed method demonstrated a linear range of 0.15 to 30 mgTM/kgfood (R2 > 0.99), with a limit of detection of 0.15 mgTM/kg food and a limit of quantification of 0.5mgTM/kgfood and successfully applied to commercially processed foods, such as potato chips, biscuits, surimi, and hot pot seasonings, which evidenced the applicability of proteomics-based methodology for food allergen analysis.

Unraveling novel umami peptides from yeast extract (Saccharomyces cerevisiae) using peptidomics and molecular interaction modeling.

Yeast extract is increasingly becoming an attractive source for unraveling novel umami peptides that are healthier and more nutritious than traditional seasonings. In the present study, a strategy for screening novel umami peptides was established using mass spectrometry-based peptidomics combined with molecular interaction modeling, emphasizing on smaller peptides than previously reported. Four representative novel umami peptides of FE, YDQ, FQEY, and SPFSQ from yeast extract (Saccharomyces cerevisiae) were identified and validated by sensory evaluation, with thresholds determined as 0.234 ± 0.045, 0.576 ± 0.175, 0.327 ± 0.057 and 0.456 ± 0.070 mmol/L, respectively. Hydrogen and ionic bonds were the main characteristic interactions between the umami peptides and the well-recognized receptor T1R1/T1R3, in which Asp 110, Thr 112, Arg 114, Arg 240, Lys 342, and Glu 264 were the key sites in ligand-receptor recognition. Our study provides accurate sequences of umami peptides and molecular interaction mechanism for the umami effect.

A high-resolution optical see-through head-mounted display with eyetracking capability.

A head-mounted display system with fully-integrated eyetracking capability offers multi-fold benefits, not only to fundamental scientific research but also to emerging applications of such technology. A key limitation of the state-of-the-art eyetracked head-mounted display (ET-HMD) technology is the lack of compactness and portability. In this paper, we present an innovative design of a high resolution optical see-through ET-HMD system based on freeform optical technology. A prototype system is demonstrated, which offers a goggle-like compact form factor, non-obstructive see-through field of view and true high-definition image resolution for the virtual display. The see-through view, via the combination of a freeform prism and corrector, achieved better than 0.5 arc minute of angular resolution for the central region of approximately 40-degrees to ensure minimal impacts on the see-through vision of an HMD user.

Sunlight-directed fluorophore-switch in photosynthesis of cyanine subcellular organelle markers for bio-imaging.

The photoconvertible fluorophore synthesis enables the light controlled imaging channels switch for accurate tracking the quantity and localization of intracellular biomolecules in chemical biology. Herein, we repurposed the photochemistry of Fischer's base and developed a sunlight-directed fluorophore-switch strategy for high-efficiency trimethine cyanine (Cy3.5/Cy3) synthesis. The unexpected sunlight-directed photoconversion of Fischer's base proceeds in conventional solvents and accelerates in chloroform via photo-oxidation and hydrogen atom transfer without using extra additives, and the heterogenous dimerization mechanism was proposed and confirmed by isolation of the reactive intermediates. The reliable strategy is employed in the photosynthesis of commercially available cytomembrane marker (DiI) and other cyanine based organelle markers with appreciable yields. Sunlight-controlled fluorophore-switch of subcellular organelle markers in living cells validated the feasibility of our strategy with cell-tolerant character. Moreover, remote control synthesis of Cy3.5 in vivo directed via sunlight further demonstrated the extended application of our strategy. Therefore, this sunlight-directed strategy will facilitate exploitation of cyanine-based probes with switched fluorescence imaging channels and further enable precise description of the dynamic variations in living cells with minimal autofluorescence and cellular disturbance.

A FRET-based ratiometric fluorescent probe for Hg2+ detection in aqueous solution and bioimaging in multiple samples.

Mercury ion, as a metal cation with great toxic effect, is widely present in various production and living environments. It seriously threatens human health and environmental safety. It is of great significance to develop convenient and effective methods for mercury ion detection. Here, we designed and synthesized a new ratiometric fluorescent probe (namely APS-NA) for the detection of mercury ions in the environment and multiple biological samples. The probe is constructed by covalently connecting two fluorophores with lipolic acid to achieve fluorescence resonance energy transfer (FRET). In the molecular structure of APS-NA, acridone is used as an energy donor, 1,8-naphthalimide is used as an energy acceptor, and a dithioacetal group is used as the reaction site for Hg2+. The intact APS-NA mainly shows the green fluorescence from the acceptor moiety 1,8-naphthalimide; the presence of Hg2+ ions would break the dithioacetal linkage between acridone and 1,8-naphthalimide; the defunctionalization of FRET would lead to bright blue fluorescence emission of acridone; thus ratiometric fluorescent detection of Hg2+ can be achieved by this recognition process. The probe not only has a large Stokes shift (Δλ = 110 nm), but also has high selectivity, high sensitivity (low detection limit 30 nM) and naked eye visualization. In addition, we have successfully used this probe for the detection Hg2+ of actual samples and imaging of a variety of organisms. These results indicate that the probe has broad application prospects.

Design and synthesis of 3,4-seco-lupane triterpene derivatives to resist myocardial ischemia-reperfusion injury by inhibiting oxidative stress-mediated mitochondrial dysfunction via the PI3K/AKT/HIF-1α axis.

In this study, 86 new seco-lupane triterpenoid derivatives were designed, synthesized, and characterized, and their protective activities against ischemia-reperfusion injury were investigated in vitro and in vivo. Structure-activity relationship studies revealed that most target compounds could protect cardiomyocytes against hypoxia/reoxygenation-induced injury in vitro, with compound 85 being the most active and exhibiting more potent protective activity than clinical first-line drugs. Furthermore, all thiophene derivatives exhibited stronger protective activity than furan, pyridine, and pyrazine derivatives, and the protective activity gradually increased with the extension of the alkyl chain and changed in the substituent. The data from the in-vitro and in-vivo experiments revealed that compound 85 protected mitochondria from damage by inhibiting excessive production of oxidative stressors, such as intracellular ROS, which in turn inhibited the apoptosis and necrotize of cardiomyocytes and reduced infarct size, thereby protecting normal cardiac function. It was associated with enhanced activation of the PI3K/AKT-mediated HIF-1α signaling pathway. Therefore, compound 85 acts as an oxidative stress inhibitor, blocks ROS production, protects mitochondria and cells from myocardial ischemia/reperfusion (MI/R) injury, and represents an effective new drug for treating MI/R injury.

Albumin nanoparticles and their folate modified counterparts for delivery of a lupine derivative to hepatocellular carcinoma.

In this study, folate polyethylene glycol CTr albumin nanoparticles (FA-PEG-CTr-NPs) targeting hepatocellular carcinoma (HCC) were prepared. The nanoparticle preparation method was optimized using single-factor and response surface analysis. The prepared nanoparticles were characterized for their particle size, zeta potential, and morphology. The particle size and zeta potential were also determined. Additionally, drug loading, encapsulation efficiency, and in vitro drug release of the nanoparticles were determined. Using the Cell Counting Kit-8 method, their cytotoxicity and their cell-targeted uptake were determined using confocal microscopy and flow cytometry. Finally, the in vivo antitumor impact and tumor-targeting ability of the nanoparticles were evaluated by determining tumor volume inhibition and drug biodistribution and performing hematoxylin-eosin (H&E) staining. It was found that CTr could be effectively encapsulated into albumin nanoparticles and functionalized. The drug loading of the two nanoparticles was 67.12 ± 2.4% and 69.33 ± 2.8%, respectively. Regarding drug release, FA-PEG-CTr-NPs (89.0%) exhibited a superior release rate to CTr-NPs (70.5%) in an acidic environment. The in vitro experiments confirmed that FA-PEG-CTr-NPs yielded better cytotoxicity and faster drug uptake results than CTr and CTr-NPs. In vivo experiments confirmed that FA-PEG-CTr-NPs exhibited markedly better tumor inhibitory activity (inhibition rate was 80.21%), drug safety, and targeting than CTr and CTr-NPs. In conclusion, functionalized nanoparticles (FA-PEG-CTr-NPs) can specifically inhibit the malignant proliferation of HCC cells and are thus a promising nanoagent for the treatment of HCC.

Immune exposure: how macrophages interact with the nucleus pulposus.

Intervertebral disc degeneration (IDD) is a primary contributor to low back pain. Immune cells play an extremely important role in modulating the progression of IDD by interacting with disc nucleus pulposus (NP) cells and extracellular matrix (ECM). Encased within the annulus fibrosus, healthy NP is an avascular and immune-privileged tissue that does not normally interact with macrophages. However, under pathological conditions in which neovascularization is established in the damaged disc, NP establishes extensive crosstalk with macrophages, leading to different outcomes depending on the different microenvironmental stimuli. M1 macrophages are a class of immune cells that are predominantly pro-inflammatory and promote inflammation and ECM degradation in the NP, creating a vicious cycle of matrix catabolism that drives IDD. In contrast, NP cells interacting with M2 macrophages promote disc tissue ECM remodeling and repair as M2 macrophages are primarily involved in anti-inflammatory cellular responses. Hence, depending on the crosstalk between NP and the type of immune cells (M1 vs. M2), the overall effects on IDD could be detrimental or regenerative. Drug or surgical treatment of IDD can modulate this crosstalk and hence the different treatment outcomes. This review comprehensively summarizes the interaction between macrophages and NP, aiming to highlight the important role of immunology in disc degeneration.

An Evaluation of Suitable Habitats for Amur Tigers (Panthera tigris altaica) in Northeastern China Based on the Random Forest Model.

Amur tigers are at the top of the food chain and play an important role in maintaining the health of forest ecosystems. Scientific and detailed assessment of the habitat quality of Amur tigers in China is the key to maintaining the forest ecosystem and also addressing the urgent need to protect and restore the wild population of Amur tigers in China. This study used the random forest method to predict the potential habitat of Amur tigers in Heilongjiang and Jilin provinces using animal occurrence sites and a variety of environmental variables. Random forests are a combination of tree predictors such that each tree depends on the values of a random vector sampled independently and with the same distribution for all trees in the forest. The generalization error for forests converges to a limit as the number of trees in the forest becomes large. The generalization error of a forest of tree classifiers depends on the strength of the individual trees in the forest and the correlation between them. The results showed that the AUC value of the test set was 0.955. The true skill statistic (TSS) value is 0.5924, indicating that the model had good prediction accuracy. Using the optimal threshold determined by the Youden index as the cutoff value, we found that the suitable habitat for Amur tigers in the field was approximately 107,600 km2, accounting for 16.3% of the total study areas. It was mainly distributed in the Sino-Russian border areas in the south of the Laoyeling Mountains at the junction of Jilin and Heilongjiang provinces, the Sino-Russian border areas of Hulin-Raohe in the eastern part of the Wanda Mountains, and the Lesser Khingan Mountain forest region. The habitat suitability of the Greater Khingan Mountain and the plain areas connecting Harbin and Changchun was relatively low. Prey potential richness was the most critical factor driving the distribution of Amur tigers. Compared with their prey, the potential habitats for Amur tigers in Heilongjiang and Jilin provinces were small in total areas, sporadically distributed, and had low continuity and a lack of connectivity between patches. This indicates that some factors may restrict the diffusion of the Amur tiger, whereas the diffusion of ungulates is less restricted. The Amur tigers in this area face a serious threat of habitat fragmentation, suggesting that habitat protection, restoration, and ecological corridor construction should be strengthened to increase population dispersal and exchange. We provide a reference for future population conservation, habitat restoration, construction of ecological migration corridors, and population exchange of Amur tigers.

[Treatment of lateral ankle joint ligament sprain by shaking and poking manipulation based on finite element method].

OBJECTIVE: To conduct a preliminary study on joint injuries of anterior and calcaneal fibular ligaments of the lateral ankle joint, and to analyze mechanism of action of shaking and poking in treating ankle joint and biomechanical properties of ankle during the recovery of joint injuries. METHODS: CT scan was performed on a male volunteer with right ankle sprain. Mimics 10.0, Solidworks 2016, Hypermesh 12.0 and Abaqus 6.13 software were used to establish 3D nonlinear finite element analysis model of foot and ankle, and the validity of model was verified. Combined with clinical study, the finite element simulation analysis was carried out on the toe flexion, dorsiflexion, varus and valgus of ankle joint under different treatment periods by adjusting elastic modulus of ligament to simulate ligament injury. RESULTS: With the treatment of shake and prick and recovery of ligament injury, the maximum stress and area with large stress on tibial pitch and fibular joint surface gradually increased under the four working conditions, and the stress value of the maximum stress ligament gradually increased, and the stress of the anterior and calcaneal fibular ligament dispersed and transferred, and the axial force gradually decreased. CONCLUSION: The finite element method was used to simulate the mechanical condition of the shaking and stamping technique, and the changes of the forces of the ligament and articular surface before and after treatment of anterior and calcaneal ligament combined injury of ankle talus were intuitively observed. The treatment effect was quantified, and could provid objective and scientific basis for clinical promotion and application of this technique.

Utilization of food waste for hydrogen-based power generation: Evidence from four cities in Ghana.

Hydrogen gas will be an essential energy carrier for global energy systems in the future. However, non-renewable sources account for 96% of the production. Food wastes have high hydrogen generation potential, which can positively influence global production and reduce greenhouse gas (GHG) emissions. The study evaluates the potential of food waste hydrogen-based power generation through biogas steam reforming and its environmental and economic impact in major Ghanaian cities. The results highlight that the annual hydrogen generation in Kumasi had the highest share of 40.73 kt, followed by Accra with 31.62 kt, while the least potential was in Tamale (3.41 kt). About 2073.38 kt was generated in all the major cities. Hydrogen output is predicted to increase from 54.61 kt in 2007 to 119.80 kt by 2030. Kumasi produced 977.54 kt of hydrogen throughout the 24-year period, followed by Accra with 759.76 kt, Secondi-Takoradi with 255.23 kt, and Tamale with 81.85 kt. According to the current study, Kumasi had the largest percentage contribution of hydrogen (47.15%), followed by Accra (36.60%), Secondi-Takoradi (12.31%), and Tamale (3.95%). The annual power generation potential in Kumasi and Accra was 73.24 GWh and 56.85 GWh. Kumasi and Accra could offset 8.19% and 6.36% of Ghana's electricity consumption. The total electricity potential of 3728.35 GWh could displace 17.37% of Ghana's power consumption. This electricity generated had a fossil diesel displacement capacity of 1125.90 ML and could reduce GHG emissions by 3060.20 kt CO2 eq. Based on the findings, the total GHG savings could offset 8.13% of Ghana's carbon emissions. The cost of power generation from hydrogen is $ 0.074/kWh with an annual positive net present value of $ 658.80 million and a benefit-to-cost ratio of 3.43. The study lays the foundation and opens policy windows for sustainable hydrogen power generation in Ghana and other African countries.

[Functional peptidomics analysis of Saccharomyces pastorianus protein hydrolysates based on different enzyme treatments].

The aim of this study is to explore differences in the peptidomics of Saccharomyces pastorianus protein hydrolysates treated with different enzymes. Briefly, differences in the peptide fingerprints and active peptides of neutral protease/papain-hydrolyzed S. pastorianus were analyzed using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) combined with PEAKS Online 1.7 analysis software, Peptide Ranker, and the BIOPEP database. Compared to traditional databases, the PEAKS Online uses de novo sequencing for analysis to obtain oligopeptides smaller than pentapeptides. It provides more comprehensive data of the peptide sample. In this study, enzymatic hydrolysates of S. pastorianus protein were prepared under the optimum conditions of neutral protease and papain respectively. In total, 7221 and 7062 polypeptides were identified in the hydrolysates of neutral protease and papain, respectively; among these polypeptides, 980 were common to the two enzymes. The 6241 and 6082 unique peptides found in the hydrolysates of neutral protease and papain, respectively, indicated that the peptide fingerprints of the two hydrolysates are quite different. Peptide Ranker predicted that 3013 (41.73%) and 3095 (43.83%) peptides were potentially bioactive in the hydrolysates of neutral protease and papain, respectively. According to the BIOPEP database, neutral protease and papain contained 295 and 357 active peptides, respectively; these peptides were mainly composed of angiotensin converting enzyme (ACE) inhibitors and dipeptidyl peptidase IV inhibitors and antioxidant peptides. The number of active peptides in the hydrolysate of papain was higher than that in the hydrolysate of neutral protease, but the total ion intensity of active peptides in the former was lower than that in the latter. This study revealed the influence of protease type on the composition of enzymatic hydrolysates from S. pastorianus protein. The above results provide a reference for the development of functional products of S. pastorianus protein peptides and the high-value utilization of yeast resources.

Profibrotic role of transcription factor SP1 in cross-talk between fibroblasts and M2 macrophages.

Fibrosis disrupts tissue balance and links to severe illnesses, impairing organ function and, in some cases, even fatality. The interaction between M2 macrophages and fibroblasts is vital for tissue equilibrium. Transforming growth factor ß1 (TGF-ß1) released by M2 macrophages plays a central role in fibrosis, regulating fibroblast activity and extracellular matrix metabolism. Targeting TGF-ß1 is key to fibrosis treatment. In our study using three fibroblast cell lines, we reveal that the M2 macrophage transcription factor SP1 enhances binding to the TGF-ß1 promoter motif, promoting TGF-ß1 transcription and activating fibroblasts (This process does not involve changes in DNA methylation levels surrounding the motif sequence). The zinc fingers in SP1's DNA-binding domain 3 are crucial for this binding. In vivo, targeting SP1 in rat ligaments significantly reduces extracellular matrix accumulation. Our findings highlight SP1 as a promising target for regulating tissue extracellular matrix and combating fibrosis.