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Background: Very few true Pseudomonas methylotrophic strains have been described, and in none of them have the pathways for one-carbon (C₁) substrate metabolism been elucidated.

Methods: The genomes of three Pseudomonas strains able to grow on methanol as the sole source of carbon (C) and energy (E) were sequenced and analyzed, and one of the strains was further characterized at the proteomic and physiological level.

Results: None of the three strains possesses a classic methanol dehydrogenase enzyme, and they apparently employ generalist type-I alcohol dehydrogenases (ADHs) to catabolize methanol to formaldehyde. In two of the strains' genomes, the only complete route encoded for incorporating methylotrophic carbon is the Calvin-Benson-Bassham (CBB) cycle, while other more typical pathways for C₁-carbon assimilation (serine cycle, ribulose monophosphate cycle) appear incomplete. The indispensability of the QedA1 alcohol dehydrogenase and of ribulose bisphosphate carboxylase for growth on methanol was demonstrated by insertion mutagenesis of the qedA1 and cbbL genes in one of the strains.

Discussion: To the author's knowledge, all wild-type methylotrophic Pseudomonadota (i.e., "Gram-negative bacteria") so far described employ a specific dehydrogenase distinctively adapted to using methanol as a substrate (MxaFI, XoxFI, or Mdh2). The methylotrophic Pseudomonas strains described here lack MDH and employ generalist ADHs, thus demoting methanol dehydrogenase (MDH) from the position of a critical enzyme for methanol utilization and expanding the range of enzymes (and genes) that enable methylotrophy in nature. The second remarkable result of this work is the discovery of the utilization of the CBB cycle by a Pseudomonas strain during methylotrophic growth, an absolute novelty for this very relevant bacterial genus.

Background: This study aimed to investigate age-related differences in anthropometric characteristics, change of direction (COD) and repeated sprint ability (RSA) performance, with and without ball control, in elite soccer players from U17, U19, and U23 categories.

Methodology: Seventy-two male players (age: 18.9 ± 2.23 years; height: 1.72 ± 0.08 m; body mass: 71.7 ± 5.04 kg; body mass index (BMI): 24.3 ± 2.61 kg/m²) from three professional soccer clubs were assessed (U17 = 24; U19 = 24; U23 = 24). After a two-month period of regular training and competition, anthropometric measures (height, body mass, body mass index) were recorded. In addition, players completed the New Multi-Change of Direction Agility Test (NMAT) and the Bangsbo RSA test, both performed with and without a ball. Testing was standardized for familiarization, warm-up, and environmental conditions.

Results: U23 players were taller and heavier than U17 and U19 players, and they showed superior COD performance without the ball compared to U17, whereas no statistically significant differences were found in COD with ball or RSA performance across age groups. Correlation analyses revealed moderate associations between anthropometric variables and COD performance (r =  - 0.35 to -0.24), while higher BMI values were related to slower agility times (r = 0.24-0.26).

Conclusions: Age-related anthropometric characteristics were associated with better COD performance without the ball, whereas COD with ball and RSA performance appear less age-dependent and more influenced by training specificity. These findings highlight the importance of incorporating technical COD drills and RSA training early in player development to align physical and technical progression.

Background: Studies have shown that vertical jump biomechanics or patterns may be related to physical fitness. This study investigated the relationship between physical fitness and drop vertical jump (DVJ) biomechanics in male college basketball players.

Methods: Health-related physical fitness was measured by 20s sit-up, core endurance and flexibility test; whilst athletic-related physical fitness by Y-balance test and dominant extremity single-leg hop distance test. Kinetics and kinematics during DVJ were evaluated by VICON.

Results: Five-level side bridge correlated negatively with angle displacement of hip adduction (p = 0.014) and positively with moment of knee flexion (p = 0.033); 8-level abdominal bridge correlated positively with moment of knee flexion (p = 0.01); ankle dorsiflexion range of motion correlated negatively with mediolateral ground reaction force (p = 0.025), angle displacement of knee flexion (p = 0.004), moment of ankle plantarflexion (p = 0.009), and positively with angle displacement of ankle plantarflexion (p = 0.015); ankle plantarflexion ROM correlated negatively with angle of knee flexion (p = 0.012); trunk flexion ROM correlated negatively with moment of ankle plantarflexion (p = 0.014).

Conclusion: Health-related physical fitness could be the alternatives for DVJ biomechanics assessment.

Background: Acute kidney injury (AKI) is a serious disease with a high incidence and easy induction. The search for innovative biomarkers and treatment methods is of great significance for improving the prognosis of patients. Autophagy is closely related to the occurrence and development of AKI. This study aims to explore the role of autophagy-related genes (ARGs) as potential biomarkers and therapeutic targets in AKI.

Methods: In this study, the gene microarray data of the GEO dataset were used to explore the molecular profile of AKI, and three machine learning algorithms were used to screen autophagy-related feature genes. To further validate the reliability of the screening results, we constructed a cisplatin-induced AKI rat model to validate potential biomarkers of machine learning screening.

Results: Machine learning analysis identified 17 differentially expressed ARGs and selected the core genes FIZ1 and FBXO21, with area under curve (AUC) values both exceeding 0.7 (95% CI [0.706-0.899]). Immune analysis revealed that the number of Mast cells resting significantly decreased in AKI samples compared to normal samples (P < 0.05). Electron microscopy observations of the cisplatin-induced AKI rat model indicated thickening of the basement membrane, fusion of foot processes, and swelling and rupture of mitochondria in the model group, suggesting a correlation between AKI and mitochondrial autophagy; Western blot results indicated a significant increase in the expression of FIZ1 and a significant decrease in FBXO21 in the AKI group (P < 0.01). The results of IHC staining were also consistent with those of Western blot results.

Conclusion: This study highlights the significant role of ARGs in AKI and identifies FIZ1 and FBXO21 as promising biomarkers with high diagnostic potential, offering new insights into the molecular mechanisms underlying AKI.

Enzymes, as key biocatalysts, are essential for advancing sustainable green technologies across diverse industrial sectors. The discovery of novel enzymes is essential for expanding their applications. In this study, we identified new lipases using an integrated screening strategy. This approach combines both structural and sequence-based methods on a large-scale metagenomic database. This strategy enabled the identification of new lipases with low sequence identity to known reference proteins. Our approach, therefore, circumvents the limitations of traditional sequence-only methods, which often fail to identify functionally similar enzymes with low sequence similarity. We first used Foldseek, a state-of-the-art structural homology search tool, to rapidly screen the database for proteins with structures similar to widely used lipases. This was followed by a rigorous sequence similarity filtering against public protein databases, yielding 711 putative novel lipases. We selected and experimentally validated three candidates, confirming their lipase activity. Further biochemical characterization revealed their notable properties including thermostability with optimal activity at 50-55 °C, and distinct alkaline activity profiles, maximal at pH of 8.0-9.0. Their unique properties, including high activity at elevated temperatures and alkaline pH, suggest potential for applications in detergent formulations, bioremediation, and industrial biocatalysis. Beyond identifying these promising enzymes, this study demonstrates the power of a combined structural and sequence-based approach for finding novel biocatalysts. This methodological innovation has broad implications for future enzyme discovery from metagenomic resources.

The transition from the Early to the Middle Jurassic was marked by significant restructuring of plesiosaur communities. While knowledge of the earliest Middle Jurassic plesiosaurs is generally limited, Toarcian plesiosaur occurrences are abundant, though the vast majority of specimens have been unearthed in the United Kingdom and Germany. Here, we reassess Lusonectes sauvagei, an early-diverging plesiosaur from the lower to middle upper Toarcian of the São Gião Formation in Portugal. Originally described as Plesiosaurus sp., it was later established as a distinct taxon closely related to taxa currently encompassed within Microcleididae. Our firsthand examination of the holotype of L. sauvagei resulted in differing interpretations of certain aspects of its morphology, prompting a detailed osteological, taxonomic, and phylogenetic reevaluation. We provide a redescription of L. sauvagei, propose a new diagnosis, and investigate its phylogenetic affinities. Although the specimen is fragmentary and poorly preserved, our study suggests that, contrary to the original interpretation, L. sauvagei is not affiliated with Microcleidus spp. The taxon remains problematic and may represent either an early-diverging pliosaurid or a plesiosauroid. Lusonectes is one of the few diagnosable plesiosaurs from the upper Lower Jurassic found outside the classic British and German localities and thus offers insights into the diversity of plesiosaurs just prior to a major event in the evolutionary history of the clade.

Background: Nontuberculous mycobacteria are opportunistic pathogens which cause infections in various tissues, with the Mycobacterium avium complex (MAC) being a major cause of pulmonary diseases. Among MAC members, Mycobacterium colombiense is a clinically significant species with recognized pathogenic potential; however, studies on its genomic structure and genetic diversity remain limited.

Methods: This study investigated the drug susceptibility profiles and performed whole-genome sequencing of 12 clinical M. colombiense isolates from the Clinical Microbiology Laboratory at Srinagarind Hospital, Khon Kaen University, Khon Kaen, Thailand.

Results: Based on minimum inhibitory concentration values, moxifloxacin resistance was most prevalent (100%), followed by linezolid (90%), clarithromycin (30%), and amikacin (30%). The presence of antibiotic resistance genes and virulence factors, including ESX secretion systems and efflux pumps, highlights the bacterium's potential for immune evasion and survival under stress. Single nucleotide polymorphism-based phylogenetic analysis revealed a close genetic relatedness between the isolates. Pangenome analysis of 16 M. colombiense genomes (12 newly sequenced and four publicly available) identified 7,771 gene clusters, comprising 4,468 core genes, 1,834 accessory genes, and 1,469 unique genes, supporting a closed pangenome structure and indicating evolutionary conservation and low genetic variability among isolates.

Conclusions: This study provides valuable insight into the genomic diversity, antimicrobial resistance profiles, and virulence potential of M. colombiense isolates. These findings enhance understanding of the pathogen and may inform clinical management, targeted diagnostic development, and therapeutic strategies.

This study was performed to compare catch abundance of three commonly used bee sampling methods within pumpkin and squash (Cucurbita pepo) fields. The three methods of sampling included visual sampling, vacuum sampling, and bowl trapping. Cucurbita pepo require bee pollinators to move pollen from male flower to female flower for fruit to set. In Cucurbita pepo the three major expected bee types that should be observed are squash bees Xenoglossa pruinosa Say (formerly Eucera (Peponapis) pruinosa) [Hymenoptera: Apidae], bumble bees (Bombus spp.) [Hymenoptera: Apidae], and honey bees (Apis mellifera L. [Hymenoptera: Apidae]). Knowing what bees are present in a grower's field could help them to make field management decisions and potentially reduce input costs, thus we chose methods where collected specimens could be submitted to experts for identification. We used visual observations as our control as we could confirm these bees were contacting flower reproductive segments, and this method is widely adopted in literature. Through this study, a total of 2,502 bees were observed, of which 82% were squash bees, 3.4% were honey bees and 2.4% were bumble bees. Bowl sampling differed significantly from visual observations for all three major bee types, having significantly fewer catches. Vacuum sampling recovered no bumble bees. Vacuum sampling was not significantly different from visual observation for honey bees but caught fewer squash bees relative to visual sampling. This study also looked at other collected morphotaxa of bees, particularly in bowl trapping, however bees beyond the three taxa presented above had limited presence in visual observations, and as such were not considered to play a major role in the pollination of these plants. This study indicates that visual observation of flowers provides a more reliable estimate of active pollinators in C. pepo fields than the two other bee sampling methods, vacuum sampling and bowl trapping, and they should likely not be deployed by growers to gain a representative sample of active pollinators in the field.

Introduction: Most of the studies on alcohol dependence (AD) emphasize its impact on the nervous system and organ functions of the human body, but its molecular mechanism is largely unknown so far. This study determines serum protein changes in alcohol-dependent patients for the identification of biomarkers and the revelation of molecular mechanisms behind alcohol dependence.

Methods: Serum samples from seven newly diagnosed alcohol-dependent patients and four healthy controls are subjected to researcher-conducted proteomic analyses using data-independent acquisition (DIA) mass spectrometry. Functional enrichment analysis of gene activity and biological pathways is performed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) to reveal key processes related to alcohol dependence. The expression of Small Nuclear Ribonucleoprotein Polypeptide B (SNRPB) is evaluated to assess its potential as a biomarker, and survival analysis is performed to investigate its clinical relevance.

Results: The mass spectrometry detected a total of 1,249 and 1,020 proteins in the serum from alcohol-dependent patients and healthy controls, respectively. Biomarker analysis identified 195 proteins as potential markers with a differentially expressed pattern. Among them, 99 proteins are upregulated and 96 proteins downregulated in alcohol-dependent patients compared with controls. The GO and KEGG pathway analysis revealed that alcohol-dependent patients exhibit enhanced activity in pathways related to adenosine triphosphate (ATP)-dependent chromatin remodeling and immune responses but reduced activity in metabolic pathways. Alcohol-dependent patients exhibited significantly higher levels of the SNRPB protein, suggesting its potential role in immune system regulation and cell growth control. Survival analysis results indicated that higher levels of SNRPB are linked to worse outcomes in liver cancer patients, specifically those with liver hepatocellular carcinoma (LIHC), suggesting it could be a useful biomarker for alcohol-related diseases.

Conclusion: This study provides valuable insights into the systemic protein changes in alcohol dependence, identifying several potential biomarkers for early diagnosis and therapeutic targeting. The upregulation of SNRPB suggests its role as a potential biomarker in clinical applications.