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Laccase (EC 1.10.3.2), as eco-friendly biocatalysts, holds immense potential for sustainable applications across various environmental and industrial sectors. Despite the growing interest, the exploration of cold-adapted laccases, especially their unique properties and applicability, remains limited. In this study, we have isolated, cloned, expressed, and purified a novel laccase from Peribacillus simplex (GenBank: PP430751), which was derived from permafrost layer. The recombinant laccase (PsLac) exhibited optimal activity at 30 °C and a pH optimum of 3.5. Remarkably, PsLac exhibited remarkable stability in the presence of organic solvents, with its enzyme activity increasing by 20 % after being incubated in a 30 % trichloromethane solution for 12 h, compared to its initial activity. Furthermore, the enzyme preserved 100 % of its activity after undergoing eight freeze-thaw cycles. Notably, the catalytic center of PsLac contains Zn2+ instead of the typically observed Cu2+ found in other laccases, and metal-ion substitution experiments raised the catalytic efficiency to 3-fold when Zn2+ was replaced with Fe2+. Additionally, PsLac has demonstrated a proficient ability to degrade phenolic pollutants, such as hydroquinone, even at a low temperature of 16 °C, positioning it as a promising candidate for environmental bioremediation and contributing to cleaner production processes.
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BACKGROUND: In this study, the relationship between C-reactive protein-albumin-lymphocyte (CALLY) index, a novel composite indicator based on inflammation and nutrition, and major adverse cardiovascular events (MACEs) was investigated in patients with ST-segment elevation myocardial infarction (STEMI). MATERIALS AND METHODS: This retrospective study included 438 patients with STEMI who were treated at a single center between January 2017 and December 2020. The CALLY index was calculated for each patient on admission. The predictive value of the CALLY index for short- and long-term MACEs was evaluated using the area under the curve (AUC) analysis, and the corresponding AUC values were calculated. Clinical characteristics were analyzed after categorizing the population based on the optimal cut-off value of the CALLY index. Multivariate Cox regression analysis was used to determine factors independently associated with MACEs, while logistic regression analysis was used to identify factors independently associated with the severity of coronary artery lesions. Kaplan-Meier estimation and log-rank test were used to assess event-free survival rates among different CALLY index groups. Additionally, Spearman's correlation test was used to determine the association between the CALLY index and the Gensini score. RESULTS: The AUC for predicting short-term MACEs in STEMI patients using the CALLY index was 0.758, while the AUC for predicting long-term MACEs was 0.740. Similarly, the AUC values were 0.815 and 0.819, respectively, when evaluating the short- and long-term mortality rates using the CALLY index. Multivariable Cox regression analysis revealed that a high CALLY index (threshold of 1.50) independently reduced the risk of short-term MACEs in patients with STEMI (hazard ratio [HR] = 0.274, 95 % confidence interval [CI] = 0.121-0.621, P=0.002). Multivariable Cox regression also demonstrated that a high CALLY index (threshold > 0.91) independently reduced the occurrence of long-term MACEs during follow-up in STEMI patients (HR=0.439, 95 % CI=0.292-0.659, P<0.001). Furthermore, multivariate logistic regression analysis revealed that a high CALLY index (threshold > 1.13) independently reduced the risk of severe coronary artery lesions in patients with STEMI (odds ratio = 0.299 [95 % CI=184-0.485], P<0.001). A positive correlation was observed between the CALLY index and the Gensini score (P<0.001). CONCLUSION: The CALLY index is a novel, convenient, and valuable prognostic indicator exhibiting a protective effect against both short- and long-term MACEs in patients with STEMI, emphasizing the significance of inflammation/nutrition in this patient population.
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Designing efficient and rapid methods for the detection of organophosphorus pesticides (OPs) residue is a prerequisite to mitigate their negative health impacts. In this study, we propose the concept of an enzyme catalysis system-based hydrogel kit integrated with a smartphone detector for in-field screening of OPs. Here, we rapidly prepared phosphotriesterase hybrid nanoflowers (PTE-HNFs) using a self-assembly strategy by adding external energy and embedded the nanocomposite in sodium alginate (SA) hydrogel to construct a target-responsive hydrogel kit. The color response of the kit is induced by catalyzing methyl parathion (MP) to produce p-nitrophenol. For on-site quantification, the color variations of the portable kit are converted into digital information through a smartphone, which exhibits an applicable linear range towards OPs. The hydrogel sensing platform demonstrates a wide linear range (1-150 µM) and low detection limit (0.15 µM) for MP while maintaining high reliability, excellent long-term stability, and ease of operation. Overall, the PTE-HNFs-based SA hydrogel kit provides a useful strategy for simple and sensitive detection of MP and holds great potential for applications in detecting OPs in food and environmental water.
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Técnicas Biosensibles , Hidrogeles , Compuestos Organofosforados , Plaguicidas , Hidrolasas de Triéster Fosfórico , Teléfono Inteligente , Plaguicidas/análisis , Técnicas Biosensibles/métodos , Hidrogeles/química , Compuestos Organofosforados/análisis , Hidrolasas de Triéster Fosfórico/química , Hidrolasas de Triéster Fosfórico/análisis , Límite de Detección , Nanocompuestos/química , Alginatos/química , Metil Paratión/análisisRESUMEN
Oligonucleotides represent a class of shorter DNA or RNA nucleic acid polymers extensively applied in the biomedical field. Despite progress in detecting and analyzing oligonucleotides, high-throughput analysis of the samples remains challenging. In this work, a high-throughput analysis method for oligonucleotide analysis was developed based on acoustic droplet ejection-open port interface-mass spectrometry (ADE-OPI-MS) technology. This approach was applied to determine the enzymatic activity of terminal deoxynucleotide transferase (TdT) for DNA synthesis, with a rate of 3 s/sample, which enhanced single-sample analysis efficiency approximately 60-fold over the previous gel analysis. After testing approximately 10,000 TdT mutants, we obtained three new variants with higher catalytic activities. Finally, by integrating these mutants, the catalytic activity of TdT was improved about 4 times compared to the starting mutant. Our results successfully established a high-throughput screening method for oligonucleotide analysis, which not only provides a foundation to engineer highly efficient TdT for ab initio synthesis of DNA but also paves the way for the potential application of oligonucleotide analysis in biomedical fields.
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Ensayos Analíticos de Alto Rendimiento , Espectrometría de Masas , Oligonucleótidos , Oligonucleótidos/química , Ensayos Analíticos de Alto Rendimiento/métodos , ADN/análisis , ADN/genética , ADN/químicaRESUMEN
Pegmolesatide, a synthetic, polyethylene-glycolylated, peptide-based erythropoiesis-stimulating agent (ESA), has been recently approved in China. Pegmolesatide is derived from the structure of endogenous erythropoietin (EPO), a natural product in mammals. This study compared the in vitro effects and selectivity of pegmolesatide to those of recombinant EPO and carbamylated EPO (CEPO) through computer-aided analyses and biological tests. The findings indicate that pegmolesatide exhibited the same stimulating effect on erythropoiesis as EPO with fewer side effects than EPO and CEPO.
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PURPOSE: To compare the short-term changes in cornea, retina, and choroid of femtosecond laser-assisted cataract surgery (FLACS) with conventional phacoemulsification (CPS) in high myopia patients with cataract. SETTING: Affiliated Hospital of Nantong University, Jiangsu Province, China. DESIGN: Prospective single-center study. METHODS: Demographics, ocular clinical features, ultrasound power, absolute phacoemulsification time, and effective phacoemulsification time were recorded for each patient. Endothelial cell density (ECD), central corneal thickness (CCT), corrected distance visual acuity (CDVA), intraocular pressure (IOP), center foveal thickness (CFT), subfoveal choroidal thickness (SFCT), and choroidal vascularity index (CVI) were evaluated preoperatively and at 1 week, 1 month, and 3 months postoperatively. Intraoperative parameters and intraoperative/postoperative complications were recorded. RESULTS: 97 eyes (46 eyes and 51 eyes in the FLACS and CPS groups, respectively) were included and analyzed. Effective phacoemulsification time was lower in the FLACS group compared with the CPS group ( P < .05). The increase in CCT was significantly lower in the FLACS group compared with the CPS group at 1 week and 1 month ( P < .05). CDVA and IOP were similar in both groups at the final visit ( P > .05). The ECD decreased was lower among CPS patients compared with FLACS patients. CFT, SFCT, and CVI increase in both groups but were increased more in the CPS group with high myopia patients. No serious complications occurred in either group. CONCLUSIONS: FLACS is a more safety and effective in cataract patients with high myopia. It has advantages in effectively reducing EPT and promoting faster recovery of the cornea, macular, and choroidal thickness.