Search Results (17,423)

The continuous increase in demand for polyethylene terephthalate (PET) has drawn global attention to the significant environmental pollution caused by the degradation of PET plastics. Exploring new PET-degrading enzymes is essential for enhancing the degradation efficiency of PET, and esterases and lipases with plastic degradation capabilities have become a focal point of research. In this study, we utilized the ultra-efficient mutant FASTase of the PET-degrading enzyme IsPETase, derived from Ideonella sakaiensis, as a positive control, based on the similarity in enzyme activity and substrate. We investigated the PET model substrate degradation activities of the esterase Gs and lipase GI, both derived from Bacillus spp., as well as the lipase CAI derived from Pseudomonas spp. The results indicated that Gs exhibited excellent bis(2-hydroxyethyl) terephthalate (BHET) degradation activity; however, Gs demonstrated a lack of thermal stability when hydrolyzing BHET. Molecular docking analyses were conducted to identify the key amino acids involved in the degradation of BHET by Gs from a structural perspective. At the same time, GI and CAI showed no BHET degradation activity. The combination of Gs and the mono-2-hydroxyethyl terephthalate (MHET) hydrolase, MHETase, can completely hydrolyze BHET, and Gs also exhibited degradation activity against the PET model substrate bis(benzyloxyethyl) terephthalate and PET nanoparticles. Given the structural similarity between PET hydrolase LCC-ICCG and Gs, this study provides new enzyme resources for advancing the efficient biological enzymatic degradation of PET plastics. Full article

High-mountain microalgae exhibit remarkable adaptability to extreme environments, making them promising candidates for sustainable biorefineries. We evaluated how photoperiod (12:12, 18:6, 24:0 h) and LED spectra (cool white, full spectrum, red–blue 4:1) affect growth and metabolite formation in Chlorella sp. UFPS019 and Scenedesmus sp. UFPS021. Biomass peaked in Chlorella under red–blue 18:6 (≈1.8 g L⁻¹) and in Scenedesmus under red–blue 24:0 (≈1.7 g L⁻¹), revealing species-specific responses. Carbohydrate fractions were maximized under red–blue 12:12 in both species, and continuous light (24:0) depressed carbohydrate content—most notably under full spectrum. Protein content was highest under red–blue 18:6 in Chlorella sp. and under red–blue 12:12–18:6 in Scenedesmus sp. Lipid fractions increased with light duration, peaking under red–blue 18:6–24:0 in Chlorella and under red–blue 18:6–24:0—with Cool White 24:0 also high—in Scenedesmus sp. Although extended illumination favored lipids, intermediate photoperiods (12:12–18:6) provided better productivity-to-energy trade-offs and broader metabolic profiles. These results show that tuning spectral composition and photoperiod to species-specific physiology enables the targeted, energy-aware production of proteins, carbohydrates, or lipids; red–blue at intermediate durations is a robust, energy-efficient regime, whereas longer exposures can be used strategically when lipid enrichment is prioritized. Full article

This study examined sarcopenia severity effects on body composition, physical performance, and mechanical properties of gait-related muscles in older women. Forty-one women aged ≥70 years participated and were classified by the following criteria: non-sarcopenia (NS, n = 15), functional sarcopenia (FS, n = 10), sarcopenia (SP, n = 9), and severe sarcopenia (SS, n = 7). Assessments included body composition, physical performance, and muscle tone, stiffness, and elasticity of the tibialis anterior (TA) and gastrocnemius medialis (GM). Group differences were analyzed using one-way ANOVA with Bonferroni post hoc tests (α = 0.05). SP and SS groups had lower body weight, BMI, appendicular skeletal muscle mass, and calf circumference compared with NS. FS demonstrated poorer physical performance than SP across all variables, with six-meter gait speed lower than SS (p < 0.05). SP exhibited significantly higher TA muscle tone, GM muscle tone and GM stiffness than NS (p < 0.05, p < 0.01, p < 0.05, respectively), while TA elasticity was significantly lower in SP (p < 0.01). These findings indicate that sarcopenia severity negatively influences body composition, muscle function, and mechanical properties, with functional sarcopenia showing the greatest impairment in performance. Early detection and targeted interventions are therefore critical to mitigate functional decline in older women. Full article

Chiral amines are vital structural motifs in pharmaceuticals and agrochemicals, where enantiomeric purity governs bioactivity and environmental behavior. We identified a novel (R)-selective amine transaminase (MwoAT) from Mycobacterium sp. via genome mining, which exhibits activity toward the synthesis of the chiral amine (R)-1-methyl-3-phenylpropylamine. The enzyme displayed optimal activity at pH 7.0 and 40 °C, with high thermostability and solvent tolerance. Using an AlphaFold3-guided semi-rational engineering strategy integrating molecular docking, alanine scanning, and saturation mutagenesis, residue L175 was pinpointed as critical for substrate binding. The resulting L175G variant exhibited a 2.1-fold increase in catalytic efficiency (k_cat/K_m) and improved thermal stability. Applied to the asymmetric synthesis of (R)-1-methyl-3-phenylpropylamine—a precursor for the antihypertensive drug dilevalol and potential scaffold for crop protection agents—the mutant achieved 26.4% conversion with ≥99.9% ee. The enzyme also accepted several ketones relevant to agrochemical synthesis, underscoring its versatility. This work delivers an engineered biocatalyst for sustainable chiral amine production and demonstrates an AI-assisted protein engineering framework applicable to both medicinal and agricultural chemistry. Full article

Background: The Tibetan Plateau, which is known for its high elevation and low oxygen levels, presents a challenging environment for its inhabitants. To adapt to these hypoxic conditions, species of Schizothoracine, a subfamily of Cyprinidae, have developed unique physiological mechanisms and functions. Transforming growth factor-β (TGF-β) is a multifunctional cytokine involved in the regulation of cell growth, differentiation, apoptosis, and the cellular immune response. However, its specific role in adaptation to hypoxia remains poorly understood. Methods: In this study, we aimed to characterize the TGF-β1 gene in Gymnocypris dobula (Gd) and Schizothorax prenanti (Sp) and to test whether TGF-β1 contributes to hypoxia adaptation in plateau Schizothoracine fish. The predicted protein for Gd-TGF-β1 contains several primary domains, including cwf21 (cdc5 protein 21), GYF (Glycine-Tyrosine-Phenylalanine), FN1 (Fibronectin 1), a conservative domain, and a signal peptide. Results: The results of tissue distribution revealed that the mRNA level of TGF-β1 in brain, heart, muscle, skin, gills, and spleen—which are key tissues involved in oxygen sensing, transport, and physiological adaptation to hypoxic environments—was significantly lower in G. dobula than that in S. prenanti. Western blotting analysis revealed that the expression of activated TGF-β1 in G. dobula was significantly higher than that in S. prenanti. To investigate whether TGF-β1 in G. dobula possesses hypoxic adaptive features, Gd-TGF-β1 and Sp-TGF-β1 were cloned into an expression vector and transfected into 293-T cells, which are widely used due to their ease of culture, high transfectability, and well-characterized properties. We found that the survival rate of cells transfected with Gd-TGF-β1 was significantly higher than that of cells transfected with Sp-TGF-β1 after hypoxia treatment. Conclusions: These findings suggest that G. dobula may promote hypoxic adaptation through the activation and increased expression of TGF-β1. Changes in TGF-β1 expression may play a role in the adaptation of G. dobula to hypoxic conditions. Full article

Probiotics like Lactobacillus sp. are extensively studied for their beneficial host interactions, including the gut–brain axis, anti-inflammatory effects, immune system interactions, restoration of gut dysbiosis, and anti-aging effects. In the current study, pearl millet was fermented with Lactobacillus plantarum strains DHCU 70 and MCC 5231, which enhanced the nutritional, bioactive, and functional properties of derived probiotic beverages. Compared to unfermented controls, fermented beverages exhibited increased protein content and vitamins B₁, B_2, and B₃, with decreased carbohydrate and dietary fiber levels. The probiotics have maintained viability exceeding 12 log CFU/mL and showed resistance to harsh gastrointestinal conditions. Fermentation increased total phenolic content from 13.38 ± 0.40 mg GAE/100 g to 42.10 ± 2.65 mg GAE/100 g (LPDB) and 47.76 ± 1.37 mg GAE/100 g (LPMB) and total flavonoid content from 13.01 ± 1.18 mg QE/100 g to 23.12 ± 2.73 mg QE/100 g and 24.21 ± 0.98 mg QE/100 g, respectively. Antioxidant assays showed DPPH radical scavenging improved by 37%, ferrous ion chelation rose from 71.69 ± 0.09 mg TE/100 g to 91.45 ± 0.006 mg TE/100 g, ABTS scavenging increased from 71.62 mg TE/100 g to 82.51 ± 0.04 mg TE/100 g (LPDB) and 89.74 ± 0.04 mg TE/100 g (LPMB) and superoxide radical inhibition rose from 51.40 ± 0.98% to 81.77 ± 0.03% (LPDB) and 79.92 ± 0.02% (LPMB). In the in vivo model, Caenorhabditis elegans, fermented beverage treatments significantly improved health-span parameters like head-swing frequency (13.51% increase), body bend frequency (8.41% increase), pharyngeal pumping (8.15% increase) with reduced lipofuscin accumulation and intracellular reactive oxygen species while median lifespan extended beyond 24 days versus 14–16 days in controls (p < 0.05). Gompertz mortality modeling revealed a significant decrease in the aging rate parameter, indicating systemic mitigation of stress-induced physiological decline. These combined nutritional, bioactive, and in vivo longevity results underscore the potential of L. plantarum-fermented pearl millet beverages as functional nutraceuticals that target oxidative stress and promote healthy aging. Full article

Transmembrane proteins (TMPs) constitute approximately 30% of the mammalian proteome and are critical targets in biomedical research due to their involvement in signaling, transport, and drug interactions. However, their unique structural characteristics pose significant challenges for conventional multiple sequence alignment (MSA) methods, which are typically optimized for soluble proteins. In this paper, we propose TMP-M2Align, a novel topology-aware multiobjective algorithm specifically designed for the multiple alignment of TMPs. The method simultaneously optimizes two complementary objectives: (i) a topology-aware Sum-of-Pairs (SPs) score that integrates region-specific substitution matrices and gap penalties, and (ii) an Aligned Regions (ARs) score that rewards consistent alignment of functional and topological domains. By combining these objectives, TMP-M2Align generates Pareto front approximations of alignment solutions, enabling researchers to select trade-offs that best suit their biological questions. We evaluated TMP-M2Align on BAliBASE Reference Set 7 and on complete datasets of human G protein-coupled receptors (GPCRs) from classes A, B1, and C. Experimental results demonstrate that TMP-M2Align consistently outperforms both traditional alignment tools and specialized TM-specific methods in terms of SPs and Total Column metrics. Moreover, qualitative topological analyses confirm that TMP-M2Align preserves the integrity of transmembrane helices and loop boundaries more effectively than competing approaches. These findings highlight the effectiveness of integrating topology-aware scoring with multiobjective optimization for achieving accurate and biologically meaningful alignments of TMPs. Full article

This observational retrospective study aimed to examine the association between team behaviour during periods of one-goal leads and subsequent match outcomes while accounting for team level and match location. All matches (n = 380) of the English Premier League (EPL) during the season 2023/24 were analyzed. Team behaviour was evaluated by shots every 10 min during a one-goal lead (SP10MDOGL), a time-normalized indicator of offensive activity that reflects a team’s strategic orientation while protecting a narrow lead. Mixed effects multinomial logistic regression was used to establish the association between SP10MDOGL and the match outcome. Results indicated that increased SP10MDOGL was strongly associated with a higher likelihood of both drawing (Odds ratio (OR) = 2.37, 95% confidence interval (CI) = 1.29–4.33; Cohen’s d (d) = 0.47) and winning (OR = 3.38; 95%CI = 1.93–5.92; d = 0.67) compared to losing. This association remained consistent across high-, intermediate-, and low-level teams regardless of whether they played at home or away. These findings suggest that maintaining an offensive approach through an increased number of shots during a one-goal lead is associated with a higher likelihood of securing positive match outcomes within the elite-level football context, such as the EPL. Soccer coaches should consider implementing proactive offensive strategies when protecting a narrow lead, regardless of their team level and match location. Full article

Objectives: To assess the incidence rate of anterior acute uveitis (AAU) in patients with longstanding axial spondyloarthritis (axSpA); to evaluate demographic and clinical characteristics associated with AAU development; and to determine the influence of AAU on bDMARD initiation and retention in this population. Methods: This two-timepoint cohort study analysed data from patients enrolled in the Spanish SpA registry REGISPONSER (2004–2007), who were re-evaluated 17 years later in the REGISPON-3 follow-up study (2021–2023). Information on the date of first AAU episode and bDMARD initiation was collected. Kaplan–Meier and Cox proportional hazards models were used to assess AAU incidence, predictors, and its association with time to bDMARD initiation and treatment retention. Results: A total of 299 patients with longstanding axSpA were included, of whom 33.4% experienced at least one episode of AAU, corresponding to an incidence rate of 1.15 per 100 person-years. The cumulative probability of a first episode of AAU increased with disease duration. The relative risk for developing a second episode after the first, compared to the overall risk of any episode in the total population, was 1.85 (95% CI: 1.34–2.57). In multivariable cox analysis, female sex and baseline enthesitis were independently associated with a higher risk of AAU. AAU did not significantly affect the likelihood of subsequent bDMARD initiation, with similar cumulative treatment probabilities in patients with and without AAU. Among treated patients, adalimumab was more frequently prescribed in those with a history of AAU. bDMARD retention rates at two and five years were comparable regardless of AAU status, suggesting that AAU was not associated with long-term treatment persistence. Conclusions: In patients with longstanding axSpA, the incidence of AAU increased steadily over time. However, the presence of AAU did not significantly influence bDMARD initiation or long-term retention in routine clinical practice. Full article

Background/Objective: Timely identification of bloodstream pathogens is critical for sepsis management; however, PCR inhibitors such as sodium polyanetholesulfonate (SPS) in blood culture broth compromise nucleic acid recovery and long read sequencing. We assessed whether coupling a benzyl alcohol SPS-removal step to the fully automated LabTurbo AIO extractor improves Oxford Nanopore-based pathogen detection. Methods: Thirteen positive blood culture broths were pre-treated with benzyl alcohol and divided: half volumes were purified on the LabTurbo AIO; paired aliquots underwent manual QIAamp extraction. DNA purity was evaluated by NanoDrop and Qubit. Barcoded libraries were sequenced on MinION R9.4.1 flow cells for 6 h. Results: Automated eluates showed a median A₂₆₀/A₂₈₀ of 1.92 and A₂₆₀/A₂₃₀ of 1.96, versus 1.80 and 1.48 for manual extracts. The automated workflow generated 1.69 × 10⁶ total reads compared with 3.9 × 10⁵ reads for manual extraction. The median N50 read length increased from 5.9 kb to 8.7 kb, and the median proportion of reads classified to species increased from 62% to 84%. The hands-on time was <5 min and the sample-to-answer turnaround was <8 h, compared with >9 h and 90 min for the manual protocol, respectively. Conclusions: Benzyl alcohol SPS removal integrated into the LabTurbo AIO extractor yielded purer, longer, and higher read counts, enhancing nanopore sequencing depth and accuracy while compressing diagnostic turnaround to a single working day. This represents a practical advance for rapid blood culture pathogen identification in critical care settings. Full article

Anaxyelidae, a relict lineage of sawflies, are represented by a single extant species today but displayed remarkable Mesozoic diversity. Here, we describe the Orthosyntexis mascula sp. nov. from mid-Cretaceous Burmese amber. The new species can be readily distinguished by its forewing, with a normally sized, uniformly sclerotized pterostigma; 1-Rs shorter than 1-M; cell 1M more than twice as long as wide; absence of 1r-rs; 1-Cu, distinctly shorter than 2-Cu; 3-Cu shorter than 4-Cu; 2m-cu shorter than 1m-cu; and 3rs-m twice as short as 4-M. In the hind wing, abscissa 2-M+Cu present, 1-M shorter than 2-M, crossvein m-cu absent, and cell R1 closed. Mesotibia with two apical spurs. Examination of high-resolution photographs of Kempendaja jacutensis enables a revised interpretation of its venation, confirming its placement in Anaxyelinae. Comparative analysis of syntexine taxa further reveals that variation in the forewing crossvein 1r-rs consistently corresponds with hind wing venation, suggesting that multiple evolutionary trajectories may have existed within Syntexinae. These findings not only expand the known diversity of Cretaceous Anaxyelidae but also provide new evidence for reconstructing the evolutionary history and internal diversification of Anaxyelidae. Full article

Optogenetics leverages light to control neural circuits, but traditional systems are often bulky and tethered, limiting their use. This work introduces OptoBrain, a novel, portable wireless system for optogenetics designed to overcome these challenges. The system integrates modules for multichannel data acquisition, smart neurostimulation, and continuous processing, with a focus on low-power and low-voltage operation. OptoBrain features up to eight neuronal acquisition channels with a low input-referred noise (e.g., 0.99 µVRMS at 250 sps with 1 V/V gain), and reliably streams data via a Bluetooth 5.0 link at a measured throughput of up to 400 kbps. Experimental results demonstrate robust performance, highlighting its potential as a simple, practical, and low-cost solution for emerging optogenetics research centers and enabling new avenues in neuroscience. Full article

Background/Objectives: The longitudinal associations between different modalities of sedentary behaviors (SBs) and sedentary patterns (SPs) with metabolic syndrome (MetS) in children and adolescents are unclear. We aimed to analyze the cross-sectional and longitudinal (2-year follow-up) association between SB and SP with the MetS score in Spanish children and adolescents. Methods: 76 children (34 females) and 186 adolescents (94 females) were included for SB analyses, and 175 children (82 females) and 188 adolescents (95 females) for SP. Children and adolescents were aged 6–11.9 years and 12–17.9 years, respectively. SB were assessed by a self-reported questionnaire and SP were determined by accelerometry. The MetS score was computed from the waist circumference, systolic blood pressure, triglycerides, high-density lipoprotein cholesterol, and glucose levels. Different linear regression models were implemented to examine cross-sectional, longitudinal, and change associations of SB and SP with MetS. Results: Total daily SB, educative daily SB, and mean SB were longitudinal and inversely associated with MetS (β = −0.001, all p < 0.05) in male adolescents, while other daily SB was longitudinal and inversely associated with MetS (β = −0.002, all p < 0.05) in female adolescents. Changes in screen and other daily SB were directly associated with MetS in female adolescents (β = 0.001 to 0.002, all p < 0.05). In contrast, changes in educative daily SB were inversely associated with MetS in female adolescents (β = −0.001, all p < 0.05). Conclusions: Few associations between SB modalities and the MetS score were found, mainly in adolescents and often in unexpected directions. In male adolescents, total and educative daily SB were negatively associated with MetS. In female adolescents, other daily SB and changes in educative daily SB showed negative associations, while changes in screen-based and other daily SB were positively associated with MetS. No associations were found between SP and MetS. Given the low evidence available to date, more longitudinal studies analyzing SB and SP simultaneously are needed to reach solid conclusions. Full article

Background/Objectives: In response to the COVID-19 pandemic, we developed a vaccine candidate against SARS-CoV-2. Spike Ferritin Nanoparticle (SpFN) comprises 24 identical prefusion-stabilized spike proteins anchored to a self-assembled nanoparticle. Organized along the three-fold axis of the ferritin particle, eight SARS-CoV-2 spike trimers are presented per nanoparticle. Methods: Here, we describe the CGMP processes for manufacturing SpFN using transient transfection of Expi293F cells. Results: The final yield of SpFN was ~10 mg per liter of media supernatant. The resulting protein is stable in cold storage for two years at −20 °C, as well as for a month at room temperature, and can withstand multiple freeze/thaw cycles. SpFN material produced using the CGMP protocols adjuvanted with Army Liposomal Formulation-QS-21 (ALFQ) elicited potent neutralizing antibodies against WA-1, Alpha, Beta, and Delta variants in mice as measured by a pseudovirus neutralization assay. Conclusions: This work demonstrates rapid development and scaled-up production of clinical-grade SARS-CoV-2 vaccine protein material, allowing permissive storage and transport conditions, and serves as a framework for recombinant protein production for future emergent pathogens. Full article

This study reports on the direct electron transfer (DET) ability of the enzyme spermidine dehydrogenase (SpDH) and its use in a DET-type enzymatic sensor for detecting spermine. SpDH was found to exhibit internal electron transfer from its cofactor, flavin adenine dinucleotide (FAD), to heme b. This was confirmed by observing the heme b-derived reduction peak at 560 nm in the presence of spermine, the substrate. SpDH was immobilized on a gold electrode via a dithiobis (succinimidyl hexanoate) self-assembled monolayer. The cyclic voltammetry analysis of the SpDH-immobilized gold electrode revealed an increased oxidation current in the presence of 0.1 mM spermine with an onset potential of −0.14 V vs. Ag/AgCl in the absence of an additional external electron acceptor. This result confirmed that SpDH is capable of DET. Chronoamperometric analyses were conducted using an SpDH-immobilized gold electrode with spermine as the substrate under a 0 V oxidation potential vs. Ag/AgCl using an artificial saliva matrix containing 10 µM ascorbic acid and 100 µM uric acid. The sensor exhibited good linear correlation between the current increase and spermine concentration from 0.2 to 2.0 µM, with a limit of detection of 0.084 µM, which encompasses the physiologically relevant spermine concentration found in the saliva. Primary structure alignments and 3D structure predictions revealed that all SpDH homologs possess two conserved histidine residues in the same location on the surface as the heme b ligand of SpDH. This indicates their potential for DET-ability with an electrode. Full article