Search Results (252)

Glutamate dehydrogenase (GDH) is ubiquitous in organisms and crucial for amino acid metabolism, energy production, and redox balance. The gdhA and gudB genes encoding GDH were identified in Bacillus altitudinis AS19 and shown to be regulated by iron. However, their functions remain unclear. In this study, gdhA and gudB were analyzed using bioinformatics tools, such as MEGA, Expasy, and SWISS-MODEL, expressed with a prokaryotic expression system, and the induction conditions were optimized to increase the yield of soluble proteins. Phylogenetic analysis revealed that GDH is evolutionarily conserved within the genus Bacillus. GdhA and GudB were identified as hydrophobic proteins, not secreted or membrane proteins. Their structures were primarily composed of irregular coils and α-helices. SWISS-MODEL predicts GdhA to be an NADP-specific GDH, whereas GudB is an NAD-specific GDH. SDS-PAGE analysis showed that GdhA was expressed as a soluble protein after induction with 0.2 mmol/L IPTG at 24 °C for 16 h. GudB was expressed as a soluble protein after induction with 0.1 mmol/L IPTG at 16 °C for 12 h. The proteins were confirmed by Western blot and mass spectrometry. The enzyme activity of recombinant GdhA was 62.7 U/mg with NADPH as the coenzyme. This study provides a foundation for uncovering the functions of two GDHs of B. altitudinis AS19. Full article

Limited molecular data exist on zoonotic parasites Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in Angus calves from Guizhou, China. This study constitutes the first molecular epidemiological survey of these pathogens in this region. 817 fecal samples from Angus calves across 7 intensive beef farms (Bijie City). Nested PCR methods targeting SSU rRNA (Cryptosporidium spp.), gp60 (Cryptosporidium bovis subtyping), bg/gdh/tpi (G. duodenalis), and ITS (E. bieneusi) coupled with DNA sequencing were employed. DNA sequences were analyzed against the NCBI. database. Statistical differences were assessed via a generalized linear mixed-effects model. Cryptosporidium spp. prevalence 23.5% (192/817; 95% CI 28.1–34.6%), with C. bovis predominating 89.6% (172/192; 95% CI 84.4–93.5%) and six subtypes (XXVIa-XXVIf). Highest infection in 4–8-week-olds 29.9% (143/479; 95% CI 25.8–34.1%) (p < 0.01). G. duodenalis: 31.3% (256/817; 95% CI 28.1–34.6%) positive, overwhelmingly assemblage E 97.6% (6/256; 95% CI 0.9–5.0%), zoonotic assemblage A was marginal 0.7% (6/817; 95% CI 0.3–1.6%). Farm-level variation exceeded 10-fold (e.g., Gantang: 55.0% (55/100; 95% CI 44.7–65.0%) vs. Tieshi: 4.9% (5/102; 95% CI 1.6–11.1%). E. bieneusi: prevalence 19.7% (161/817; 95% CI 17.0–22.6%), exclusively zoonotic genotypes BEB4: 49.7% (80/161; 95% CI 41.7–57.7%); I: 40.4% (65/161; 95% CI 32.7–48.4%). Strong diarrhea association (p < 0.01) and site-specific patterns (e.g., Guanyindong: 39.2%). While Giardia exhibited the highest prevalence (31.3%) with minimal zoonotic risk, Enterocytozoon—despite lower prevalence (19.7%)—posed the greatest public health threat due to exclusive circulation of human-pathogenic genotypes (BEB4/I) and significant diarrhea association, highlighting divergent control priorities for these enteric parasites in Guizhou calves. Management/Public health impact: Dominant zoonotic E. bieneusi genotypes (BEB4/I) necessitate: 1. Targeted treatment of 4–8-week-old Angus calves. 2. Manure biofermentation (≥55 °C, 3 days), and 3. UV-disinfection (≥1 mJ/cm²) for karst water to disrupt transmission in this high-humidity region. Full article

In recent years, early olive fruit drop has been observed in the northern Mediterranean regions, causing significant economic losses, although the exact cause remains unknown. Recent studies have identified several possible causes; however, our understanding of how olive trees respond to these environmental stresses remains limited. This study includes an analysis of selected meteorological and flowering data for Olea europaea L. “Istrska belica” to evaluate the use of a chilling and forcing model for a better understanding of flowering time dynamics under a changing climate. The flowering process is influenced by high diurnal temperature ranges (DTRs) during the pre-flowering period, resulting in earlier flowering. Despite annual fluctuations due to various climatic factors, an increase in DTRs has been observed in recent decades, although the mechanisms by which olive trees respond to high DTRs remain unclear. The chilling requirements are still well met in the region (1500 ± 250 chilling units), although their total has declined over the years. According to the Chilling Hours Model, chilling units—referred to as chilling hours—represent the number of hours with temperatures between 0 and 7.2 °C, accumulated throughout the winter season. Growing degree hours (GDHs) are strongly correlated with the onset of flowering. These results suggest that global warming is already affecting the synchrony between olive tree phenology and environmental conditions in the northern Mediterranean and may be one of the reason for the green drop. Full article

Nitrogen (N) critically regulates wheat growth and grain quality, yet the molecular mechanisms underlying foliar nitrogen application remain unclear. This study evaluated the effects of foliar nitrogen application (12.25 kg ha⁻¹) on the growth, grain yield, and quality of spring wheat, as well as its molecular mechanisms. The results indicated that N was absorbed within 3 h post-application, with leaf nitrogen concentration peaking at 12 h. The N treatment increased whole-plant dry matter accumulation and grain protein content by 11.34% and 6.8%, respectively. Amino acid content peaked 24 h post-application, increasing by 25.3% compared to the control. RNA-sequencing analysis identified 4559 and 3455 differentially expressed genes at 3 h and 24 h after urea treatment, respectively, these DEGs being primarily involved in nitrogen metabolism, photosynthetic carbon fixation, amino acid biosynthesis, antioxidant systems, and nucleotide biosynthesis. Notably, the plastidic glutamine synthetase gene (GS2) is crucial in the initial phase of urea application (3 h post-treatment). The pronounced downregulation of GS2 initiates a reconfiguration of nitrogen assimilation pathways. This downregulation impedes glutamine synthesis, resulting in a transient accumulation of free ammonia. In response to ammonia toxicity, the leaves promptly activate the GDH (glutamate dehydrogenase) pathway to facilitate the temporary translocation of ammonium. This compensatory mechanism suggests that GS2 downregulation may be a key switch that redirects nitrogen metabolism from the GS/GOGAT cycle to the GDH bypass. Additionally, the upregulation of the purine and pyrimidine metabolic routes channels nitrogen resources towards nucleic acid synthesis, and thereby supporting growth. Amino acids are then transported to the seeds, culminating in enhanced seed protein content. This research elucidates the molecular mechanisms underlying the foliar response to urea application, offering significant insights for further investigation. Full article

Background/Objectives: Early childhood anemia is a severe public health concern and the most common blood disorder worldwide, especially in emerging countries. This study examines the sources of childhood anemia in Ghana through various societal, parental, and child characteristics. Methods: This research used data from the 2022 Ghana Demographic and Health Survey (GDHS-2022), which comprised 9353 children. Using STATA 13 and R 4.4.2 software, we analyzed maternal, social, and child factors using a model-building procedure, logistic regression analysis, and machine learning (ML) algorithms. The analyses comprised machine learning methods including decision trees, K-nearest neighbor (KNN), logistic regression, and random forest (RF). We used discrimination and calibration parameters to evaluate the performance of each machine learning algorithm. Results: Key predictors of childhood anemia are the father’s education, socioeconomic status, iron intake during pregnancy, the mother’s education, and the baby’s postnatal checkup within two months. With accuracy (94.74%), sensitivity (82.5%), specificity (50.78%), and AUC (86.62%), the random forest model was proven to be the most effective machine learning predictive model. The logistic regression model appeared second with accuracy (67.35%), sensitivity (76.16%), specificity (56.05%), and AUC (72.47%). Conclusions: Machine learning can accurately predict childhood anemia based on child and paternal characteristics. Focused interventions to enhance maternal health, parental education, and family economic status could reduce the prevalence of early childhood anemia and improve long-term pediatric health in Ghana. Early intervention and identifying high-risk youngsters may be made easier with the application of machine learning techniques, which will eventually lead to a healthier generation in the future. Full article

Long-term monoculture of poplar plantations for industrial material production has been widely reported to cause severe soil degradation, while the presence of understory vegetation might enhance soil nitrogen (N) transformation and supply. This study employed a field experiment using a randomized block design with three blocks and four understory treatments, including understory removal, N-fixing species planting, single-species retention, and diverse vegetation retention, in poplar plantations on a mid-latitude alluvial plain in China over 6 years to assess the effects of different species and richness of understory on soil N transformation and related microbial traits via ¹⁵N assays and shotgun metagenomics. The results showed that understory removal significantly reduced soil N transformation rates, bacterial abundance, and gene abundance associated with N transformation. Compared to a single-species understory, retaining a diverse understory with high species richness significantly increased soil gross N transformation rate of mineralization by 149%, nitrification by 221%, and immobilization by 85%; comprehensively enriched dominant bacterial phyla; and elevated gene abundances of gdh_K15371, ureB, hao, and amoA_B associated with N transformation. No significant difference in N transformation rates existed between N-fixing species planting treatment and single-species retention treatment, while N-fixing species planting treatment specifically promoted the soil bacterial phyla Nitrospirae and Chloroflexi, and increased the gene abundances of gdh_K15371 and hao. These findings demonstrate that both introducing N-fixing species and an increase in species richness of the understory effectively promoted soil N transformation but that different underlying mechanisms existed. Planting N-fixing species selectively increased the soil bacterial phyla of Nitrospirae and Chloroflexi, whereas the increase in species richness broadly enriched soil bacterial diversity, thereby inducing the enrichment of the functional genes and enhancing soil N transformation. In conclusion, both planting N-fixing species and retaining diverse understory vegetation were effective strategies for maintaining sustainable management of poplar plantations by increasing soil N availability. Full article

Clostridioides difficile infection (CDI), the most common cause of nosocomial diarrhea, presents with a wide spectrum of clinical manifestations, ranging from mild diarrhea to severe, life-threatening conditions such as pseudomembranous colitis and toxic megacolon. In recent years, both the incidence and severity of CDI have increased, leading to a significant burden in terms of morbidity, mortality, and healthcare costs. We conducted a single-center, retrospective cohort study for 30 months at “Sf. Parascheva” Infectious Diseases Clinical Hospital Iași, in North Eastern Romania, aiming to assess the clinical and laboratory characteristics of CDI, as well as treatment approaches and their association with patient outcomes. A total of 534 patients were included during the study period, of whom 484 had favorable outcomes, while 50 have died of the disease. Fever (p = 0.007) and age over 65 (p = 0.001) were associated with prolonged hospitalization. Patients positive for both A and B toxins and GDH had the highest risk of recurrence (p = 0.020). Among comorbidities, obesity was the only condition significantly linked to recurrence (p = 0.001). In female patients over 65 years old, the probability of survival drops below 60% after 21 days of hospitalization, highlighting a critical risk factor in this population. These results underscore the importance of comprehensive risk assessment in CDI, particularly focusing on advanced age and comorbidities, to guide early therapeutic interventions, optimize patient management, and improve clinical outcomes among high-risk populations. Full article

Backgroud: Glutamate dehydrogenase (GDH) is involved in the metabolism of glutamate and ammonia. It is regulated by multiple ligand variants, and hyper-active GDH mutants have been reported for hyperinsulinism hyperammonemia syndrome (HHS). Methods: Here, we constructed the wild-type human GDH and three human GDH454 mutants and investigated their degradation activity and performance under different GDH inhibitors. Results: Protein activity test and SDS-PAGE analysis of the purified proteins showed that the GDH454 mutant from HHS has weaker GDH enzymatic activity but greater resistance to trypsin hydrolysis than the wild type. Interestingly, using the biomolecular interactions technique, it showed that the GDH454 mutant has 10⁹ times weaker affinity for trypsin and 10-fold weaker for epigallocatechin gallate (EGCG) than the wild-type GDH. Subsequently, native-PAGE gel analysis demonstrated that EGCG could break down the GDH hexamer into monomers and form a complex with trypsin to enhance the degradation of both types of GDH. Conclusions: EGCG showed good affinity to both the wild-type and the mutant GDH proteins, promoting protein degradation; this provides a new strategy for the treatment of HHS and other hyper-active GDH-related diseases. Full article

Cryptosporidium spp. and Giardia intestinalis are important zoonotic protozoa that are closely related to diarrhea and cause considerable economic losses in the livestock breeding industry. Ganzi is one of the main production areas for yaks in China, but there have been few reports on the occurrence of Cryptosporidium spp. and G. intestinalis in yaks. This study used PCR-based sequencing techniques to survey the prevalence and species/genotypes of Cryptosporidium spp. and G. intestinalis in faecal samples from 223 yaks in Ganzi, Sichuan Province. The positive rate of Cryptosporidium spp. was 7.2% (16/223), with the highest positive rate of yaks found in Yajiang (37%, 10/27), which was significantly higher than that in Litang (3.1%, 4/130) and Seda (3%, 2/66). The positive rate of Cryptosporidium spp. in young animals aged <6 months (20.5%, 8/39) was significantly higher than that in older animals aged 12–23 months (0; 0/43) and >24 months (3.3%, 3/90). Three Cryptosporidium species were found by sequence analysis of 18S rRNA locus, namely C. andersoni, C. ryanae, and C. bovis. The total positive rate of G. intestinalis was 15.7% (35/223), with significant differences identified between Yajiang (40.7%, 11/27), Litang (17.7%, 23/130), and Seda (1.5%, 1/66). One genotype (assemblage E) was found by analyzing the sequence of gdh, bg, and tpi loci. Meanwhile, co-infection of Cryptosporidium spp. and Giardia intestinalis was identified in five samples. The present study explores the infection of Cryptosporidium spp. and G. intestinalis from yaks in Ganzi, aiming to enrich our understanding of the occurrence of these protozoa in livestock. Full article

This study aims to improve the operational efficiency of district heating (DH) systems by introducing a novel control method based on variable flow rate control, without compromising indoor comfort. The novelty of this work lies in its integrated analysis of flow control and substation configurations in DH networks, linking real-world operational strategies with mathematical modeling to improve energy efficiency and infrastructure costs. Using a case study from Omsk, Russia, where supply temperatures and energy demand profiles are traditionally rigid, the proposed approach utilizes operational data, including outdoor temperature, supply/return temperature, and hourly consumption patterns, to optimize heat delivery. A combination of flow rate adjustments, bypass line implementation, and selective control strategies for transitional seasons (fall and spring) was modeled and analyzed. The methodology integrates heat meter data, indoor temperature tracking, and Supervisory Control and Data Acquisition (SCADA)-like system inputs to dynamically adapt supply temperatures while avoiding overheating and reducing distribution losses. The results show a significant reduction in excess heat supply during warm days, with improvements in heat demand prediction accuracy (17.3% average error) compared to standard models. Notably, the optimized configuration led to a 21% reduction in total greenhouse gas (GHG) emissions (including 6537 tons of CO₂ annually), a 55.3% decrease in annualized operational costs, and a positive net present value (NPV) by year nine, with an internal rate of return (IRR) of 25.4%. Compared to conventional scenarios, the proposed solution offers better economic performance without requiring extensive infrastructure upgrades. These findings demonstrate that flexible, data-driven DH control is a feasible and sustainable alternative for aging networks in cold-climate regions. Full article

Infections with zoonotic pathogens have received increasing attention in recent years, as reflected in the literature of both veterinary and human medicine. Cryptosporidium and Giardia are recognised as the principal causes of waterborne outbreaks worldwide, but there is still limited data on the role of wild carnivores, such as red foxes and wolves, as reservoir hosts and in disseminating these pathogens in the environment. The aim of the current project was to analyse the prevalence and abundance of Cryptosporidium and Giardia infections in foxes from seven voivodeships and in wolves from the Warmia-Masuria Voivodeship in Poland and to conduct a phylogenetic analysis of the detected parasites. For the detection of both parasites, we used the commercial immunofluorescent assay MeriFluor Cryptosporidium/Giardia. For Cryptosporidium detection we also applied modified Ziehl–Neelsen (ZN) staining of faecal smears and, following PCR amplification, sequenced the 18S rDNA locus. For Giardia detection, we sequenced the glutamate dehydrogenase (gdh) gene. In total, 117 and 69 faecal samples obtained from red foxes and wolves, respectively, were screened for the presence of Cryptoporidium/Giardia. In red foxes, prevalence was 38.5% and 15.4% for Cryptosporidium spp. and G. intestinalis, respectively. In wolves, the prevalence of Cryptosporidium spp. was 14.5%, and only one sample was Giardia-positive. Cryptosporidium canis, Cryptosporidium sp. vole genotype, C. baileyi and Cryptosporidium sp. were identified in red foxes, while C. canis and Cryptosporidium sp. were detected in wolves. Our results indicate that red foxes and grey wolves act as reservoir hosts of Cryptosporidium spp. and G. intestinalis in natural areas in Poland. Full article

Effectively utilizing aquatic plants to absorb nitrogen from water bodies and convert it into organic nitrogen via nitrogen assimilation enzyme activity reduces water nitrogen concentrations. This serves as a critical strategy for mitigating agricultural non-point source pollution in the Yellow River Basin However, emergent plants’ rate and mechanism of uptake of different forms of nitrogen remain unclear. This study determined the nitrogen uptake rates, nitrogen assimilation activities, root properties, and photosynthetic parameters of four emergent plants, Phragmites australis, Typha orientalis, Scirpus validus, and Lythrum salicaria, under five NH₄⁺/NO₃⁻ ratios (9:1, 7:3, 5:5, 3:7, and 1:9) using ¹⁵N hydroponic simulations. The results demonstrated that both the form of nitrogen and the plant species significantly influenced the nitrogen uptake rates of emergent plants. In water bodies with varying NH₄⁺/NO₃⁻ ratios, P. australis and T. orientalis exhibited significantly higher inorganic nitrogen uptake rates than S. validus and L. salicaria, increasing by 11.83–114.69% and 14.07–130.46%, respectively. When the ratio of NH₄⁺/NO₃⁻ in the water body was 9:1, the uptake rate of inorganic nitrogen by P. australis reached its peak, which was 729.20 μg·N·g⁻¹·h⁻¹ DW (Dry Weight). When the ratio of NH₄⁺/NO₃⁻ was 5:5, the uptake rate of T. orientalis was the highest, reaching 763.71 μg·N·g⁻¹·h⁻¹ DW. The plants’ preferences for different forms of nitrogen exhibited significant environmental plasticity. At an NH₄⁺/NO₃⁻ ratio of 5:5, P. australis and T. orientalis preferred NO₃⁻-N, whereas S. validus and L. salicaria favored NH₄⁺-N. The uptake rate of NH₄⁺-N by the four plants was significantly positively correlated with glutamine synthetase and glutamate synthase activities, while the uptake rate of NO₃⁻-N was significantly positively correlated with NR activity. These findings indicate that the nitrogen uptake and assimilation processes of these four plant species involve synergistic mechanisms of environmental adaptation and physiological regulation, enabling more effective utilization of different nitrogen forms in water. Additionally, the uptake rate of NH₄⁺-N by P. australis and T. orientalis was significantly positively correlated with glutamate dehydrogenase (GDH), suggesting that they are better adapted to eutrophication via the GDH pathway. The specific root surface area plays a crucial role in regulating the nitrogen uptake rates of plants. The amount of nitrogen uptake exerted the greatest total impact on the nitrogen uptake rate, followed by root traits and nitrogen assimilation enzymes. Therefore, there were significant interspecific differences in the uptake rates of and physiological response mechanisms of emergent plants to various nitrogen forms. It is recommended to prioritize the use of highly adaptable emergent plants such as P. australis and T. orientalis in the Yellow River irrigation area. Full article

Background: Neisseria meningitidis (N. meningitidis) is a leading cause of acute meningitis and is classified into 13 serogroups, six of which are predominantly associated with invasive meningococcal disease. This study aimed to investigate the genotype, subgenotype, and antigenic profiles of N. meningitidis serogroup B strains isolated in Vietnam. Methods: Genotyping was performed on 106 N. meningitidis strains isolated from clinical samples from Vietnamese patients and nasopharyngeal swabs of healthy adolescents between 2019 and 2024. The genetic profiles, including the porA, porB, fetA, fHbp, abcZ, adk, aroE, fumC, gdh, pdhC, and pgm genes, were analyzed using Sanger sequencing and bioinformatic methods. Results: We found that 84.9% of the strains carried VR3 families 36 or 35-1, with VR1, VR2, and VR3 families 22-25, 14, and 36 being the most prevalent. Among the 106 serogroup B isolates, 20 variants of the porB allele 3 were identified, with porB 3-1212 being the most frequent (30.2%). Dominant PorB variable loops included L1.6, L4.5, L5.7, L6.6, and L7.13. fHbp variant group 2 was predominant (104/106 strains), and 12 FetA allele variants were identified, with F1-7 being the most common (47.2%). Three clonal complexes were identified, and clonal complex ST-32 was the most predominant. Fifty-five strains (51.9%) belonged to sequence types that have not yet been assigned to any clonal complexes, and 15 strains (14.1%) with allelic profiles were not assigned to STs. The 3-253 and 3-1212 alleles of porB, the F1-7 variant of FetA, the ST-44 and ST-1576 sequence types, and the ST-41/44 complex were observed more frequently in patients compared to asymptomatic carriers, suggesting their association with more virulence. Conclusions: This study showed a high genetic and antigenic diversity of N. meningitidis serogroup B isolates in Vietnam, with VR3 family 36 most common and porB 3-1212 as the predominant allele. fHbp variant group 2 and FetA allele F1-7 were most frequent. ST-32 was the dominant clonal complex, though many strains remained unassigned, highlighting the need for ongoing molecular surveillance. Full article

This study investigated the effects of dietary carbohydrate levels (control 8.13%, HG1 12.03%, and HG2 14.15%) on growth performance and glutamate metabolism in Chinese perch (S. chuatsi) (initial weight: 39.12 ± 0.25 g) reared at 12–15 °C. Diets were isonitrogenous (49% protein). After 8 weeks, the HG1 group optimized weight gain rate (WGR), specific growth rate (SGR), and protein efficiency ratio (PER), while reducing feed conversion ratio (FCR). HG1 and HG2 groups reduced liver glutamate/glutamine levels while downregulating the expression of key ammonia-metabolizing genes (gs, gdh, and ampd), collectively suppressing glutamate-mediated ammonia excretion. HG1 and HG2 groups enhanced glycolysis (upregulated gk and pk) coupled with suppressed gluconeogenesis (decreased PEPCK and G6Pase activities) in the liver. Concurrent downregulation of proteolytic markers (mafbx and murf1) in the muscle indicated improved protein conservation efficiency in the HG1 and HG2 groups. The HG1 diet optimally enhances growth by promoting glycolysis, reducing ammonia excretion, and improving feed efficiency. The insights gained from this research will be used to refine the low-temperature culture feed for Chinese perch, aiming to decrease ammonia and nitrogen emissions, thereby advancing the practice of low-ammonia emission culture for this species. Full article

This study aimed to better understand the impacts of various nitrogen (N) forms on blueberry growth and development, as well as to increase blueberry (Vaccinium spp.) N utilization efficiency. We selected the blueberry cultivar ‘Anna’ as the experimental material, and four N treatments were applied throughout the key vegetative growth stage: N deficiency (CK), ammonium-N (T1), nitrate-N (T2), and amide-N (T3). The growth parameters, physiology indexes, and ultrastructure changes in blueberry seedlings were explored. At the same time, the Pearson correlation model was used to analyze the correlation among each physiology index. The results showed that blueberry plants grew better under T1 and T3 treatments, with increased biomass, N content, chlorophyll content, and photosynthetic efficiency. Under T1 treatment, the leaves had lower O₂^˙− generation rate and MDA concentration, but higher superoxide dismutase (SOD), glutamate synthetase (GOGAT), and glutamine synthetase (GS) activity. Compared to T1 treatment, T2 treatment dramatically enhanced peroxidase (POD) activity, glucose content, and free amino acid content, particularly Arg content. Furthermore, N deficit treatment inhibited plant growth while increasing free radicals, POD, catalase (CAT), and glutamate dehydrogenase (GDH) activities, as well as the content of antioxidant compounds. Correlation and principal component analysis showed that photosynthetic properties, chlorophyll content, antioxidant system, amino acid levels, and N metabolizing enzyme activity were significantly affected by different N forms. This study can serve as a scientific foundation for optimal N regulation and management in blueberries. Full article