Metabolomic discoveries for early diagnosis and traditional Chinese medicine efficacy in ischemic stroke.

Ischemic stroke (IS), a devastating cerebrovascular accident, presents with high mortality and morbidity. Following IS onset, a cascade of pathological changes, including excitotoxicity, inflammatory damage, and blood-brain barrier disruption, significantly impacts prognosis. However, current clinical practices struggle with early diagnosis and identifying these alterations. Metabolomics, a powerful tool in systems biology, offers a promising avenue for uncovering early diagnostic biomarkers for IS. By analyzing dynamic metabolic profiles, metabolomics can not only aid in identifying early IS biomarkers but also evaluate Traditional Chinese Medicine (TCM) efficacy and explore its mechanisms of action in IS treatment. Animal studies demonstrate that TCM interventions modulate specific metabolite levels, potentially reflecting their therapeutic effects. Identifying relevant metabolites in cerebral ischemia patients holds immense potential for early diagnosis and improved outcomes. This review focuses on recent metabolomic discoveries of potential early diagnostic biomarkers for IS. We explore variations in metabolites observed across different ages, genders, disease severity, and stages. Additionally, the review examines how specific TCM extracts influence IS development through metabolic changes, potentially revealing their mechanisms of action. Finally, we emphasize the importance of integrating metabolomics with other omics approaches for a comprehensive understanding of IS pathophysiology and TCM efficacy, paving the way for precision medicine in IS management.

Novel pH- and thermal-responsive oleogel capsules: Featuring an oleogel core and ultrathin calcium-alginate shell.

Oleogels have been explored as a new lipid-based delivery system, however, their insolubility and unsuitable shape severely limit their application in food systems. Herein, core-shell oleogel capsules with high monodispersity (coefficient variation (CV) < 5%)) were prepared via gravity-assisted co-flowing microfluidic device and simply air-drying. The oleogel capsules with oleogel core and ultrathin calcium-alginate shell were prepared. Oleogel capsules maintained their original shape at pH = 2.0 but swelled rapidly at pH = 6.8 and 7.4. The swelling ratio of shell can be adjusted by inner fluid flow rate (Qin). Notably, the core with beeswax (BW) crystal network, effectively improved the stability performances and also could provide thermal response. Finally, the oleogel capsules demonstrated excellent sustained release and UV protection of lipophilic bioactives. This work sheds light on development of novel oleogel capsules, making them ideal candidates for smart food encapsulation applications.

Metal-Free Electron Donor-Acceptor Pair Enabled Long-Term Stability of Li-CO2 Battery.

The challenges of Lithium-carbon dioxide (Li-CO2) batteries for ensuring long-term cycling stability arise from the thermodynamically stable and electrically insulating discharge products (e.g., Li2CO3), which primarily rely on their interaction with the active materials. To achieve the optimized intermediates, the bifunctional electron donor-acceptor (D-A) pairs are proposed in cathode design to adjust such interactions in the case of B-O pairs. The inclusion of BC2O sites allows for the optimized redistribution of electrons via p-π conjugation. The as-obtained DO-AB pairs endow the enhanced interactions with Li+, CO2, and various intermediates, accompanied by the adjustable growth mode of Li2CO3. The shift from solvation-mediated mode into surface absorption mode in turn manipulates the morphology and decomposition kinetics of Li2CO3. Therefore, the corresponding Li-CO2 battery got twofold improved in both the capacity and reversibility. The cycling prolongs exceed 1300 h and well operates at a wide temperature range (20-50 °C) and different folding angles (0-180°). Such a strategy of introducing electron donor-acceptor pairs provides a distinct direction to optimize the lifetime of Li-CO2 battery from local structure regulation at the atomic scale, further inspiring in-depth understandings for developing electrochemical energy storage and carbon capture technologies.

Relationship between biofilm formation and antibiotic resistance of Klebsiella pneumoniae and updates on antibiofilm therapeutic strategies.

Klebsiella pneumoniae is a Gram-negative bacterium within the Enterobacteriaceae family that can cause multiple systemic infections, such as respiratory, blood, liver abscesses and urinary systems. Antibiotic resistance is a global health threat and K. pneumoniae warrants special attention due to its resistance to most modern day antibiotics. Biofilm formation is a critical obstruction that enhances the antibiotic resistance of K. pneumoniae. However, knowledge on the molecular mechanisms of biofilm formation and its relation with antibiotic resistance in K. pneumoniae is limited. Understanding the molecular mechanisms of biofilm formation and its correlation with antibiotic resistance is crucial for providing insight for the design of new drugs to control and treat biofilm-related infections. In this review, we summarize recent advances in genes contributing to the biofilm formation of K. pneumoniae, new progress on the relationship between biofilm formation and antibiotic resistance, and new therapeutic strategies targeting biofilms. Finally, we discuss future research directions that target biofilm formation and antibiotic resistance of this priority pathogen.

UBD participates in neutrophilic asthma by promoting the activation of IL-17 signaling.

Neutrophilic asthma is a persistent and severe inflammatory lung disease characterized by neutrophil activation and the mechanisms of which are not completely elucidated. Ubiquitin D (UBD) is a ubiquitin-like modifier participating in infections, immune responses, and tumorigenesis, while whether UBD involves in neutrophilic asthma needs further study. In this study, we initially found that UBD expression was significantly elevated and interleukin 17 (IL-17) signaling was enriched in the endobronchial biopsies of severe asthma along with neutrophils increasing by bioinformatics analysis. We further confirmed that UBD was upregulated in the lung tissues of neutrophilic asthma mouse model. UBD overexpression promoted IL-17 signaling activation. Knockdown of UBD suppressed the activation of IL-17 signaling. UBD interacted with TRAF2 and reduced the total and the K48-linked ubiquitination of TRAF2. However, IL-17 A stimulation increased both the total and the K48-linked ubiquitination of TRAF2. Together, these findings indicated that UBD was upregulated and played a critical role in IL-17 signaling which contributed to a better understanding of the complex mechanisms in neutrophilic asthma.

Exploring the potential of VGLL3 methylation as a prognostic indicator for intracranial aneurysm with gender-specific considerations.

DNA methylation is widely recognized to play a role in intracranial aneurysm (IA) pathogenesis. We investigated the levels of methylation of vestigial-like 3 (VGLL3) in IA and explored its potential as a prognostic indicator. A total of 48 patients with IA and 48 healthy controls were included in the present study. Methylation levels of CpG sites were assessed using bisulfite pyrosequencing, and levels of VGLL3, TEAD, and YAP in the blood were measured by real-time quantitative polymerase chain reaction testing. VGLL3 methylation was significantly higher in controls than in IA patients (P=0.001), and this phenomenon was more pronounced in females (P<0.001). Compared with the control group, the expression levels of VGLL3 and TEAD in the blood of IA patients were significantly increased, while YAP was significantly decreased. VGLL3 methylation was positively correlated with HDL (P=0.003) and female Lpa concentration (r = 0.426, P=0.03), and was also negatively correlated with age (P=0.003), APOE (P=0.005), and VGLL3 mRNA expression (P<0.001). Methylation and mRNA expression of VGLL3 may serve as indicators of IA risk in females (AUC = 0.810 and 0.809). VGLL3 methylation may participate in the pathogenesis of IA by regulating the expression of the VGLL3/TEAD/YAP pathway, and its gene methylation and expression levels have IA risk prediction value.

Guard Cell-Inspired Ion Channels: Harnessing the Photomechanical Effect via Supramolecular Assembly of Cross-Linked Azobenzene/Polymers.

Stimuli-responsive ion nanochannels have attracted considerable attention in various fields because of their remote controllability of ionic transportation. For photoresponsive ion nanochannels, however, achieving precise regulation of ion conductivity is still challenging, primarily due to the difficulty of programmable structural changes in confined environments. Moreover, the relationship between noncontact photo-stimulation in nanoscale and light-induced ion conductivity has not been well understood. In this work, a versatile design for fabricating guard cell-inspired photoswitchable ion channels is presented by infiltrating azobenzene-cross-linked polymer (AAZO-PDAC) into nanoporous anodic aluminum oxide (AAO) membranes. The azobenzene-cross-linked polymer is formed by azobenzene chromophore (AAZO)-cross-linked poly(diallyldimethylammonium chloride) (PDAC) with electrostatic interactions. Under UV irradiation, the trans-AAZO isomerizes to the cis-AAZO, causing the volume compression of the polymer network, whereas, in darkness, the cis-AAZO reverts to the trans-AAZO, leading to the recovery of the structure. Consequently, the resultant nanopore sizes can be manipulated by the photomechanical effect of the AAZO-PDAC polymers. By adding ionic liquids, the ion conductivity of the light-driven ion nanochannels can be controlled with good repeatability and fast responses (within seconds) in multiple cycles. The ion channels have promising potential in the applications of biomimetic materials, sensors, and biomedical sciences.

An efficient and universal In silico screening strategy for acquisition of high-affinity Aptamer and its application in analytical utility.

Numerous aptamers against various targets have been identified through the technology of systematic evolution of ligands by exponential enrichment (SELEX), but the affinity of these aptamers are often insufficient due to the limitations of SELEX. Therefore, a more rational in silico screening strategy (ISS) was developed for efficient screening of high affinity aptamers, which took shape complementarity and thermodynamic stability into consideration. Neuron specific enolase (NSE), a tumor marker, was selected as the target molecule. In the screening process, three aptamer candidates with good shape complementarity, lower ΔG values, and higher ZDOCK scores were produced. The dissociation constant (Kd) of these candidates to NSE was determined to be 10.13 nM, 14.82 nM, and 2.76 nM, respectively. Each of them exhibited higher affinity to NSE than the parent aptamer (Kd = 23.83 nM). Finally, an antibody-free fluorescence aptasensor assay, based on the aptamer with the highest affinity, P-5C8G, was conducted, resulting in a limit of detection (LOD) value of 1.8 nM, which was much lower than the parental aptamer (P, LOD = 12.6 nM). The proposed ISS approach provided an efficient and universal strategy to improve the aptamer to have a high affinity and good analytical utility.

No causal effect of genetically determined circulating homocysteine levels on psoriasis in the European population: evidence from a Mendelian randomization study.

Background: Although numerous studies demonstrated a link between plasma homocysteine (Hcy) levels and psoriasis, there still exists a certain level of controversy. Therefore, we conducted a Mendelian randomization study to investigate whether homocysteine plays a causative role in the development or exacerbation of psoriasis. Methods: A two-sample Mendelian randomization (MR) analysis was conducted. Summary-level data for psoriasis were acquired from the latest R9 release results from the FinnGen consortium (9,267 cases and 364,071 controls). Single nucleotide polymorphisms (SNPs) robustly linked with plasma Hcy levels at the genome-wide significance threshold (p < 5 × 10-8) (18 SNPs) were recognized from the genome-wide meta-analysis on total Hcy concentrations (n = 44,147 participants) in individuals of European ancestry. MR analyses were performed utilizing the random-effect inverse variance-weighted (IVW), weighted median, and MR-Egger regression methods to estimate the associations between the ultimately filtrated SNPs and psoriasis. Sensitivity analyses were conducted to evaluate heterogeneity and pleiotropy. Results: MR analyses revealed no causal effects of plasma Hcy levels on psoriasis [IVW: odds ratio (OR) = 0.995 (0.863-1.146), p = 0.941; weighed median method: OR = 0.985 (0.834-1.164), p = 0.862; MR-Egger regression method: OR = 0.959 (0.704-1.305), p = 0.795]. The sensitivity analyses displayed no evidence of heterogeneity and directional pleiotropy, and the causal estimates of Hcy levels were not influenced by any individual SNP. Conclusion: Our study findings did not demonstrate a causal effect of genetically determined circulating Hcy levels on psoriasis.

Peroxymonocarbonate activation via Co nanoparticles confined in metal-organic frameworks for efficient antibiotic degradation in different actual water matrices.

Traditional advanced oxidation processes suffer from low availability of ultrashort lifetime radicals and declining stability of catalysts. Co nanoparticles in hollow bimetallic metal-organic frameworks (Co@MOFs) were synthesized via a solvothermal method. Nanoconfinement and peroxymonocarbonate (PMC) degradation system endows Co@MOFs with high catalytic activity and stability even in the actual water matrices. The nanocomposites exhibited 100-200 nm polyhedron structure with irregular nanocavity between the 20 nm shell and multicores. Co nanoparticles were completely encapsulated by the FeIII-MOF-5 shell according to the X-ray diffraction and photoelectron spectra. Both 0.8 nm micropores and 3.6 nm mesopores were proven to be present. The yolk-shell Co@MOFs exhibited higher catalytic performance than that of Co nanoparticles, hollow FeIII-MOF-5 and its core-shell counterpart toward PMC activation during sulfamethoxazole degradation. The catalytic activities of Co@MOFs for the activation of unsymmetrical peroxides (PMC and peroxymonosulfate) were much higher than those for the symmetrical peroxides (H2O2 and persulfate) and the heterogeneous catalysis was dominant in the Co@MOFs activated H2O2 and PMC systems. The MOF stability was the highest and metal leakages were the least in the activated PMC system among the four peroxides because of mild reaction conditions and the alkalescent solution (pH = 8.3-8.4). Furthermore, the high removal efficiencies (>94%) and degradation rates could be maintained in the different actual water matrices due to the confinement effects. The contributions of carbonate and hydroxyl radicals were primary for sulfamethoxazole degradation, and superoxide anion and singlet oxygen also played essential roles according to scavenging experiments and time-series spin-trapping electron spin resonance spectra. Six degradation pathways were proposed according to 26 intermediate identification and the pharmacophores of more than 80% intermediates were destroyed, which would benefit subsequent biological treatment. Successful combination of nanoconfinement and PMC might provide a new effective solution for pollution remediation.

Association between serum albumin-to-creatinine ratio and clinical outcomes among patients with ST-elevation myocardial infarction after percutaneous coronary intervention: a secondary analysis based on Dryad databases.

Background: The prognostic value of the serum albumin-to-creatinine ratio (sACR) in patients with ST-elevation myocardial infarction (STEMI) remains unclear. This study aims to investigate the impact of the sACR on incident major adverse cardiovascular events (MACEs) among revascularized patients with STEMI at long-term follow-up. Methods: A total of 461 patients with STEMI who underwent successful primary percutaneous coronary intervention (PCI) were enrolled to explore the association between the sACR and MACE during a 30-month follow-up. The Cox regression proportional hazard model was used to evaluate the prognostic value of the sACR. Heterogeneity among specific groups was investigated by subgroup analysis. Results: A total of 118 patients developed MACE during the follow-up. A negative association between the sACR and MACE was found after adjusting for other MACE-related risk factors. In subgroup analyses, the sACR was inversely associated with MACE in patients aged ≥ 60 years [hazard ratio (HR), 0.478; 95% confidence interval (CI), 0.292-0.784], male (HR, 0.528; 95% CI, 0.327-0.851), with hypertension history (HR, 0.470; 95% CI, 0.271-0.816), and with anterior wall myocardial infarction (HR, 0.418; 95% CI, 0.239-0.730). Meanwhile, the negative association between the sACR and MACE remained significant in a sensitivity analysis that excluded patients with low serum albumin levels (HR, 0.553; 95% CI, 0.356-0.860). Conclusions: Patients with STEMI who underwent successful PCI with a low sACR had a higher risk of developing MACE, indicating that the sACR could be used to identify patients with STEMI who are at high risk of developing MACE.

Application of intraoperative infrared thermography in bypass surgery for adult moyamoya syndrome: A preliminary study.

Background and objectives: Cerebral revascularization surgery is the mainstay of treatment for moyamoya syndrome (MMS) today, and intraoperative determination of the patency of the revascularized vessel is a critical factor in the success of the procedure. Currently, major imaging modalities include intraoperative indocyanine green (ICG) videoangiography (ICG-VA), digital subtraction angiography (DSA), and vascular ultrasound Doppler. Infrared thermography is a modern imaging modality with non-contact devices for the acquisition and analysis of thermal data. We aimed to investigate the feasibility and advantages of infrared thermography in determining anastomotic patency during MMS surgery. Methods: Indocyanine green videoangiography and infrared thermography were performed simultaneously in 21 patients with MMS who underwent bypass surgery. The detection result of vessel patency was compared, and the feasibility and advantages of infrared thermography were assessed. Results: The patency of the anastomosis was accurately determined in 21 patients using either ICG angiography or infrared thermography. In 20 patients, the results of infrared thermography showed that the vascular anastomosis was unobstructed, and there was an agreement with the subsequent results of ICG-VA. In one patient, we suspected inadequate patency after testing the anastomosis with infrared thermography, and the results of ICG-VA evaluation of the anastomosis confirmed that there was indeed an anastomotic obstruction. Conclusion: Compared with ICG-VA, infrared thermography might offer an alternative non-invasive, contrast-free option in assessing anastomosis patency compared with ICG-VA, and it is likely to become more widely used in the clinic in the near future.

Precise electronic structure modulation on MXene-based single atom catalysts for high-performance electrocatalytic CO2 reduction reaction: A first-principle study.

Electrocatalytic CO2 reduction reaction (CO2RR) to CO is a logical approach to achieve a carbon-neutral cycle. In this work, a series of Ti2CO2 and O vacancy containing Ti2CO2 MXene-based transition metal (TM) single atom catalysts (SACs), including TM-Ti2CO2 and TM-Ov-Ti2CO2, are explored for high-performance CO2RR. Sc/Ti/V/Cr-Ti2CO2 and Ni-Ov-Ti2CO2 are screened out with limiting potential (UL) more positive than -0.50 V. Ni-Ov-Ti2CO2 is a candidate catalyst for CO2RR to CO, considering its activity with UL of -0.27 eV, and the selectivity relevant to hydrogen evolution reaction and HCOOH production. Meanwhile, a novel activity descriptor of TM-Ti-O group valence state is proposed according to that TMs work in synergy with coordinated Ti and O atoms and a level of around 0.64 e- benefits to CO2RR. This work highlights oxygen vacancy containing Ti2CO2-based Ni SAC as a promising catalyst for CO2RR, and provides a feasible electronic structure design principle for guiding the design of MXene-based SACs for CO2RR.

Highly-efficient peroxydisulfate activation by polyaniline-polypyrrole copolymers derived pyrolytic carbon for 2,4-dichlorophenol removal in water: Coupling mechanism of singlet oxygen and electron transfer.

Carbonization of N-containing aromatic polymers is a promising route to prepare N-doped carbon materials with low cost, easy regulation, and no external N source. However, there are relatively few studies applying these materials for persulfate activation, and the catalytic mechanisms of the existing reaction systems are divergent. In this paper, a series of N-doped carbon materials were prepared by carbonizing polyaniline (PANI), polypyrrole (PPy), and PANI-PPy copolymers. The copolymer-derived carbon materials exhibit superior peroxydisulfate (PDS) catalytic activity compared to some commercially available and reported carbon materials. Combing quenching experiments, EPR analysis, chemical probe analysis, and various electrochemical analysis methods identified the singlet oxygen (1O2) and electron transfer as the main reaction pathways of all systems, but the contribution of each pathway was influenced by the types of precursors. The structure-activity relationship indicated that the carbonyl group (CO) was the main active site for the 1O2 pathway, while the electron transfer ability of the reaction system and the potential of the complex formed by catalyst and PDS jointly determined the electron transfer pathway. This paper provides a new strategy for obtaining excellent N-doped carbon-based persulfate activators and deepens the insight into the mechanism of PDS activation by N-doped carbon materials.

Gene Repair of iPSC Line with GARS (G294R) Mutation of CMT2D Disease by CRISPR/Cas9.

OBJECTIVE: Charcot-Marie-Tooth disease (CMT) severely affects patient activity, and may cause disability. However, no clinical treatment is available to reverse the disease course. The combination of CRISPR/Cas9 and iPSCs may have therapeutic potential against nervous diseases, such as CMT. METHODS: In the present study, the skin fibroblasts of CMT type 2D (CMT2D) patients with the c.880G>A heterozygous nucleotide mutation in the GARS gene were reprogrammed into iPSCs using three plasmids (pCXLE-hSK, pCXLE-hUL and pCXLE-hOCT3/4-shp5-F). Then, CRISPR/Cas9 technology was used to repair the mutated gene sites at the iPSC level. RESULTS: An iPSC line derived from the GARS (G294R) family with fibular atrophy was successfully induced, and the mutated gene loci were repaired at the iPSC level using CRISPR/Cas9 technology. These findings lay the foundation for future research on drug screening and cell therapy. CONCLUSION: iPSCs can differentiate into different cell types, and originate from autologous cells. Therefore, they are promising for the development of autologous cell therapies for degenerative diseases. The combination of CRISPR/Cas9 and iPSCs may open a new avenue for the treatment of nervous diseases, such as CMT.

Screening and verification of genes related to polycystic ovary syndrome.

OBJECTIVE: To identify key genes involved in occurrence and development of polycystic ovary syndrome (PCOS). METHODS: By downloading the GSE85932 dataset from the GEO database, we used bioinformatical analysis to analyse differentially expressed genes (DEGs) from blood samples of eight women with PCOS and eight matched controls. Following bioinformatic analysis, we performed a cross-sectional study of serum samples taken from 79 women with PCOS and 36 healthy controls. RESULTS: From the 178 DEGs identified by bioinformatical analysis, 15 genes were identified as significant, and of these, ORM1 and ORM2 were selected for further verification as potential biomarkers for PCOS. Serum ORM1 and ORM2 levels were significantly increased in women with PCOS, and had a high diagnostic value. ORM1 and ORM2 were positively correlated with testosterone, cholesterol, and triglycerides. ORM1 levels were negatively correlated with high density lipoprotein (HDL) while ORM2 levels showed no significant correlation. CONCLUSIONS: ORM may be an effective biomarker for the diagnosis of PCOS and its monitoring may be a useful therapeutic strategy.

Quercetin Mitigates Radiation-Induced Intestinal Injury and Promotes Intestinal Regeneration via Nrf2-Mediated Antioxidant Pathway1.

Radiation-induced intestinal injury is one the most common adverse events of radiotherapy, which can severely affect quality of life. There are currently no effective preventive and therapeutic options for this disorder. Quercetin is a natural flavonoid found in common food species, with the characteristics of antioxidative, anti-inflammatory, and anti-cancerous activity. However, the role of quercetin on radiation-induced intestinal injury and the underlying mechanism remains poorly understood. In this study, we found quercetin treatment can improve the survival rate of mice after a single-dose (10 Gy) abdominal irradiation. Quercetin-pretreated mice significantly reduced radiation-induced DNA damage and intestinal epithelium cell apoptosis. In addition, quercetin also improved the proliferation activity of intestinal stem cells and promoted intestine epithelium repair after irradiation. Further studies demonstrated that quercetin treatment curtailed radiation-induced reactive oxygen species generation via regulating Nrf2 signaling in intestinal epithelium cells. Furthermore, treatment with Nrf2 inhibitor, could reverse the above effects. Altogether, quercetin can ameliorate radiation-induced intestine injury via regulating Nrf2 signaling, scavenging free radicals, and promoting intestinal epithelium repair.

Photoswitchable and Solvent-Controlled Directional Actuators: Supramolecular Assembly and Crosslinked Polymers.

Untethered small actuators have drawn tremendous interest owing to their reversibility, flexibility, and widespread applications in various fields. For polymer actuators, however, it is still challenging to achieve programmable structural changes under different stimuli caused by the intractability and single-stimulus responses of most polymer materials. Herein, multi-stimuli-responsive polymer actuators that can respond to light and solvent via structural changes are developed. The actuators are based on bilayer films of polydimethylsiloxane (PDMS) and azobenzene chromophore (AAZO)-crosslinked poly(diallyldimethylammonium chloride) (PDAC). Upon UV light irradiation, the AAZO undergoes trans-cis-trans photoisomerization, causing the bending of the bilayer films. When the UV light is off, a shape recovery toward an opposite direction occurs spontaneously. The reversible deformation can be repeated at least 20 cycles. Upon solvent vapor annealing, one of the bilayer films can be selectively swollen, causing the bending of the bilayer films with the directions controlled by the solvent vapors. The effects of different parameters, such as the weight ratios of AAZO and film thicknesses, on the bending angles and curvatures of the polymer films are also analyzed. The results demonstrate that multi-stimuli-responsive actuators with fast responses and high reproducibility can be fulfilled.

Genetic Arg-304-His substitution in GRK5 protects against sepsis progression by alleviating NF-κB-mediated inflammation.

BACKGROUND: Previous studies have demonstrated that G protein-coupled receptor kinase 5 (GRK5) exerts a pivotal regulatory effect on the inflammation associated with sepsis. The present study aimed to investigate the clinical association of GRK5 genetic variants with sepsis and to further explore the underlying genetic mechanisms involved in regulating sepsis-induced inflammatory responses and the pathogenesis of sepsis. METHODS: This case-control study enrolled 1081 septic patients and 1147 matched controls for genotyping of GRK5 rs2230349 and rs2230345 polymorphisms. The effect of these genetic variants on GRK5-mediated inflammatory responses was analyzed in peripheral blood mononuclear cells (PBMCs) and THP-1 macrophages. A clinically relevant polymicrobial sepsis model was established by subjecting wild-type (WT) and GRK5-knockout mice to cecal ligation and puncture (CLP) to evaluate the role of GRK5 in sepsis. RESULTS: We identified significant differences in the genotype/allele distribution of rs2230349 G > A, but not rs2230345, between the sepsis subtype and septic shock subgroups (GA + AA vs. GG genotype, OR = 0.698, 95% CI = 0.547-0.893, P = 0.004; A vs. G allele, OR = 0.753, 95% CI = 0.620-0.919, P = 0.005) and between the survivor and nonsurvivor subgroups (GA + AA vs. GG genotype, OR = 0.702, 95% CI = 0.531-0.929, P = 0.015; A vs. G allele, OR = 0.753, 95% CI = 0.298-0.949, P = 0.017). PBMCs carrying the sepsis-associated protective A allele produced significantly lower levels of TNF-α and IL-1ß upon LPS stimulation. The results from the in vitro experiment showed that the Arg-304-His substitution caused by the rs2230349 G-to-A mutation in GRK5 significantly decreased the LPS-induced production of several proinflammatory cytokines, such as TNF-α, IL-6, IL-1ß and MCP-1, via the IκB-α/NF-κB signaling pathway in THP-1 macrophages. Furthermore, GRK5-knockout mice exhibited a significant decrease in IκB-α phosphorylation/degradation, the p-p65/p65 ratio, the p-p50/p50 ratio, p65 nuclear translocation and downstream cytokine (TNF-α, IL-6, IL-1ß and VCAM-1) production compared to WT mice after CLP surgery. A significant improvement in 7-day survival rate in GRK5-KO septic mice was observed in the presence of antibiotics. CONCLUSIONS: The Arg-304-His substitution caused by the rs2230349 G-to-A mutation in GRK5 might disrupt GRK5 function and alleviate IKB-α/NF-κB-mediated inflammatory responses, which ultimately conferred a genetic protective effect against susceptibility to sepsis progression and mortality. These results may, to some extent, explain the heterogeneity of the clinical prognoses of septic patients and provide novel opportunities for individualized approaches for sepsis treatment.

Characterization of a novel detergent-resistant type I pullulanase from Bacillus megaterium Y103 and its application in laundry detergent.

This study aims to find a moderate pullulanase for detergent industry. The pulY103B gene (2217 bp) from Bacillus megaterium Y103 was cloned and expressed in Escherichia coli. PulY103B contained four conserved regions of glycoside hydrolase family (GH) 13 and the typical sequence of type I pullulanase. The optimal reaction conditions of PulY103B were pH 6.5 and 40 °C. In addition, it remained stable below 40 °C and over 80% of activity was retained at pH ranging from 6.0 to 8.5. The best substrate for the enzyme was pullulan. Furthermore, it exhibited activity toward wheat starch (36.5%) and soluble starch (33.4%) but had no activity toward amylose and glycogen. Maltotriose and maltohexaose were major pullulan hydrolysis products. Soluble starch and amylopectin were mainly hydrolyzed into maltotetraose. These results indicated that PulY103B is a novel type I pullulanase with transglycosylation activity via formation of α-1,4-glucosidic linkages. Moreover, PulY103B was strongly stimulated by nonionic detergents [viz, Tween 20 (10%), Tween 80 (1%), Triton X-100 (20%)] and commercial liquid detergents (3.0 g/L). Wash performance tests demonstrated that the mixture of PulY103B and detergent removed starch-based stains better than using detergent alone (p < 0.05). Therefore, this pullulanase has big potential as a detergent additive.