Digitized imaging of the spatial distribution of a targeted metal over a given area converts a continuous spectrum of data into a set of discrete pixels of intensities that ideally correlate strongly with the areal density map. This should be independent of the metal imaging modality, allowing semiquantitative analysis. In practice, correlation strength may vary between data sets obtained by different modalities. One enduring problem for all modalities is selecting a cutoff threshold intensity appropriately distinguishing the true signal from background noise. This may entail subjective choices between "conservative" thresholds prioritizing specificity over sensitivity (i.e., true positives despite possible false negatives) and "discovery-driven" thresholds prioritizing identification of putative effects for subsequent validation (i.e., true negatives despite possible false positives). Computerized data processing may help make this more objective by comparing outcomes for the set of all possible threshold values, termed cumulative threshold analysis. We address pitfalls in performing valid cumulative threshold analysis using ImageJ/Fiji for relative quantification of brain iron in mice with normal or genetically elevated iron, assessed by classical histochemistry or synchrotron X-ray fluorescence. In addition to pitfalls in choosing settings for generating analysis histograms, these include data loss with conversion between bit depths and selecting both appropriate X-axis directionality and image display range minima and maxima for analyzing multiple images. If these factors are handled appropriately, cumulative thresholding provides a powerful approach for more objective analysis of biometal imaging. This has important applications for metal imaging in research and clinical settings.
The presence of naturally occurring radionuclides in coastal groundwater constitutes a significant public health risk, especially in areas susceptible to hydrological variability. This research applies multivariate statistical methods, including principal component analysis and hierarchical cluster analysis, alongside the PHREEQC thermodynamic model to elucidate the mechanisms governing the mobility of naturally occurring radioactive materials (NORM), specifically gross alpha and beta activity, in a tropical coastal aquifer. Examination of 44 groundwater samples collected over two seasons indicated that more than half exceeded the national guideline for gross alpha activity (0.1 Bq/L). The results indicate that mildly acidic and reducing (anaerobic) conditions facilitate the reductive dissolution of iron and manganese hydroxides. The reductive dissolution of Fe/Mn oxides destabilizes the mineral structures that previously acted as sorptive barriers, thereby releasing trapped radionuclides directly into the groundwater. Additionally, this mobilization is synergistically influenced by competitive ion exchange mechanisms. PHREEQC modeling further identifies a potential iron "lock/release" mechanism: at mildly acidic conditions (mean pH 5.36-5.39), ferrihydrite precipitation is inhibited, maintaining elevated radionuclide mobility. Conversely, as pH increases (up to 7.32), the system approaches supersaturation (saturation index, SI > 0), promoting co-precipitation and resulting in a more than 31-fold decrease in alpha activity during the rainy season (from 0.405 to 0.013 Bq/L). In contrast, beta-emitting radionuclides are less affected by iron-related processes and are predominantly influenced by dilution, with a reduction of approximately threefold (from 0.358 to 0.109 Bq/L). These findings establish a scientific basis for developing cost-effective water treatment approaches, such as aeration combined with pH adjustment, to simultaneously remove heavy metals and alpha radioactivity from tropical coastal aquifer systems.
Green leafy vegetables (GLVs) are vital components of a healthy diet and contribute substantially to dietary trace element intake. This study evaluated trace element concentrations in GLVs grown under conventional, organic, and hydroponic farming systems (n = 27) and marketed in Jaipur, Rajasthan, India. A total of 12 elements (Ba, Be, B, Co, Li, Mn, Mo, Se, Ag, Ti, V, and Zn) were analyzed using ICP-MS. Kruskal-Wallis H and Mann-Whitney U test were used for inferential statistical analyses depending on vegetable type and farming system. Principal component analysis (PCA) was used to identify principal components representing major sources of variation. Hydroponically grown samples exhibited higher concentrations of key essential trace elements, particularly B (12.36 mg/kg), Mn (14.29 mg/kg), and Zn (13.92 mg/kg), than conventional (4.99, 10.48, and 4.84 mg/kg) and organic samples (2.54, 7.68, and 5.20 mg/kg) respectively. Organic samples generally showed lower trace element levels, with negligible concentrations of Be, Co, Li, Se, and V. Conventional samples displayed intermediate but more variable elemental profiles. The results for Kruskal-Wallis analysis indicated significant differences (p < 0.05) for most elements except Mn and Ti. Pairwise comparisons confirmed significant differences between hydroponic and organic systems for several elements (p < 0.001). PCA revealed clear separation among farming systems. Overall, the findings highlight that farming practices influence the elemental composition of GLVs and are useful for evaluating their nutritional quality.
Rice genotypes exhibit differential iron-toxicity tolerance, where tolerant types maintain iron homeostasis through plaque formation, sequestration and antioxidant defense, while sensitive genotypes accumulate excess iron, causing oxidative damage and cellular dysfunction. Iron (Fe) toxicity is a major constraint to rice production in waterlogged acidic soils, where excessive accumulation of ferrous iron disrupts cellular homeostasis and limits crop productivity. Indigenous rice germplasm represents a valuable source of adaptive traits for improving stress tolerance; however, the mechanisms underlying Fe toxicity tolerance in traditional rice cultivars from Northeast India remain poorly understood. In this study, 55 indigenous rice genotypes from Assam were evaluated under hydroponic iron stress to identify physiological, biochemical, and molecular traits associated with tolerance. Considerable genotypic variation was observed in germination, seedling growth, biomass retention, root and shoot tolerance indices, and leaf bronzing symptoms. Tolerant genotypes, including Ahom Sali, Phul Pakhori, Prasad Bhog, and Mahsuri, maintained higher biomass, lower leaf bronzing, enhanced root iron plaque formation, restricted iron translocation to shoots, and greater antioxidant enzyme activities than susceptible genotypes. These responses were associated with reduced oxidative damage and improved maintenance of nutrient homeostasis under excess iron conditions. Gene expression analysis performed in one representative tolerant genotype (Ahom Sali) and one susceptible genotype (Ranjit Sub-1) revealed differential regulation of OsFer1, OsVIT2, OsIRT1, and OsFeSOD2, providing preliminary evidence that iron sequestration and antioxidant defense contribute to Fe toxicity tolerance. Overall, the study demonstrates that coordinated regulation of root iron sequestration, intracellular iron storage, and antioxidant defense underpins Fe toxicity tolerance in indigenous rice. The identified tolerant genotypes represent valuable genetic resources for developing Fe toxicity-resilient rice cultivars adapted to acid soil ecosystems.
Lymphatic filariasis (LF) is a neglected tropical disease causing chronic lymphatic dysfunction and disability. Selenium, an essential trace element with antioxidant and immunomodulatory functions, may influence disease progression, yet its status in patients with filarial lymphedema remains understudied. This study aimed to assess serum levels in patients affected with filarial lymphedema. A case-control study was conducted at the Filariasis Management Clinic of ICMR-Vector Control Research Centre, Puducherry, including 40 filarial lymphedema patients and 30 healthy controls. Sociodemographic data, anthropometric measurements, and dietary information were collected through semi-structured interviews. Serum selenium concentrations were measured using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in an NABL-accredited laboratory. Statistical analysis employed Fisher's exact test for categorical variables and Student's t-test for continuous variables. Patients demonstrated significantly higher selenium deficiency prevalence (47.5.0% vs 16.7%, p<0.001) compared to controls, representing 3-fold elevated risk. Despite adequate macronutrient intake, micronutrient deficiency persisted across all lymphedema grades (35% Grade 2, 27.5% Grade 3, 37.5% Grade 4). Vitamin supplementation showed protective effects against selenium deficiency (p=0.004). This pilot study highlights significant selenium deficiency among patients affected with filarial lymphedema. These findings suggest potential benefits of integrating selenium screening and targeted nutritional interventions into comprehensive lymphatic filariasis management programs. Further longitudinal studies are warranted to establish causal relationships and evaluate therapeutic interventions.
This study aimed to assess the relationship between hepcidin, IFN-γ, iron biomarkers, and clinical variables in patients with anemia of inflammation. A cross-sectional study was conducted. Ninety patients with inflammatory anemia and 70 healthy controls were evaluated. Laboratory parameters, serum hepcidin, and IFN-γ were determined using ELISA. The mean age of patients with anemia of inflammation was 74.1±7.8 years, and most were female. Most patients reported ≥3 comorbidities (70%). Patients with ≥3 comorbidities had significantly higher levels of hepcidin (p<0.001). Hemoglobin, transferrin saturation index, and serum iron levels were significantly lower in patients with anemia of inflammation than in the control group (p<0.001). The erythrocyte sedimentation rate and ferritin and hepcidin levels increased in the anemia group (p<0.05). A significant positive correlation was observed between hepcidin and ferritin (r=0.372; p=0.001) and IFN-γ (r=0.228; p=0.004). Conversely, a significant negative correlation was found between hepcidin and hemoglobin (r=-0.355; p<0.001). Understanding how inflammatory parameters and upregulated hepcidin levels act as biomarkers to assess iron homeostasis in anemia of inflammation may be effective for differential diagnosis and therapeutic approaches, such as iron supplementation.
Trace elements play essential roles in neurobiology, but evidence linking whole blood concentrations to cognitive outcomes in the general population remains limited. This study examined whether concentrations of eight trace elements: selenium, mercury, cadmium, arsenic, lead, manganese, copper, and zinc were associated with cognitive performance assessed approximately six years later in a Norwegian general population cohort from the HUNT3 Survey. Whole blood trace element concentrations were measured using high resolution inductively coupled plasma mass spectrometry, with several strategies to minimize contamination. General cognitive performance was assessed using a validated web-based cognitive test battery capturing processing speed, motor speed, attention, and episodic and working memory. Associations were estimated using linear models with restricted cubic splines, adjusting for age, sex, and education. The sample included 252 participants aged 50-65 years (43% women), generally characterized by good somatic and mental health. No associations were observed for selenium, mercury, cadmium, arsenic, lead, manganese, or copper. Zinc showed a shallow U-shaped association, with slightly lower cognitive performance at mid-range concentrations, although the effect size was small. Cognitive performance declined with age and increased with educational attainment. In this low-exposure general population, whole blood trace elements concentrations showed no meaningful associations with cognitive performance six years later. These findings suggest that exposure levels to non-essential trace elements in this cohort are too low to exert clinically relevant effects on cognitive health. Alternatively, single time-point whole blood measurements may not reflect cumulative exposure relevant to long-term cognitive outcomes, or the study was underpowered to detect modest associations.
Emerging evidence highlights a link between heavy metal exposure and semen quality, but discrepancies persist regarding the effects of metals like lead and cadmium across different biological matrices. This systematic review and meta-analysis aimed to explore the association between heavy metal exposure in blood, urine, and seminal plasma and male semen quality. A comprehensive search was conducted across four databases (PubMed, Embase, Scopus, and Cochrane Library), and a bibliometric analysis was performed using the Web of Science Core Collection. Based on this analysis, a systematic literature review and meta-analyses were conducted. Database searches were updated until November 2024. A total of 10 qualitative and 14 quantitative studies were included. Exposure to lead and cadmium in seminal plasma was significantly correlated with progressive sperm motility, with odds ratio (OR) of 2.27 (95% confidence interval [CI]: 1.18 to 4.34) for lead and of 2.55 (95% CI: 1.29 to 5.03) for cadmium. Arsenic exposure was significantly linked to total sperm motility (OR = 2.88, 95% CI: 1.35 to 6.17), while zinc deficiency in seminal plasma was associated with reduced total sperm motility (OR = 1.77, 95% CI: 1.17 to 2.66). Given the widespread exposure to heavy metals, assessing their impact on male reproductive health should be prioritized as a public health concern. This study highlights the reprotoxicity of heavy metals in seminal plasma, blood, and urine, with seminal plasma emerging as the most sensitive biomarker for reproductive risk.
Kashin-Beck disease (KBD) is a chronic, endemic osteoarticular disorder associated with T-2 toxin exposure, which is rapidly metabolized to HT-2 toxin in vivo. However, the role of zinc transporter ZIP6 in HT-2 toxin-induced extracellular matrix metabolic disturbance in chondrocytes remains unclear. This study established HT-2 toxin intervention and ZIP6 knockdown chondrocyte models, combined with quantitative reverse transcription polymerase chain reaction (qRT-PCR) and transcriptome sequencing, to investigate the regulatory mechanisms of ZIP6 in chondrocyte injury. Following HT-2 toxin exposure, altered chondrocyte morphology and reduced cell viability were observed; ZIP6, COL2A1, MTF1, and MTF2 expression were significantly downregulated, whereas MMP1, MMP13, and COL10A1 were upregulated. ZIP6 knockdown significantly upregulated COL2A1, MTF1, and MTF2, downregulated MMP1 and COL10A1, but did not alter MMP13 expression. Differentially expressed genes following ZIP6 knockdown were predominantly enriched in FoxO, cAMP, and TNF signaling pathways. qRT-PCR validation confirmed consistent expression changes in FOXO4, TNFSF10, COLEC10, UCA1, and LRRC17 with transcriptome sequencing results. Collectively, HT-2 toxin suppresses ZIP6, MTF1, and MTF2 expression, disrupting chondrocyte extracellular matrix metabolism; conversely, ZIP6 knockdown ameliorates matrix metabolic abnormalities through modulation of relevant signaling pathways. These findings reveal differential regulatory effects of HT-2 toxin and ZIP6 on cartilage matrix metabolism, providing experimental evidence for elucidating molecular mechanisms underlying cartilage injury-related diseases.
This randomized clinical trial evaluated the ability to negotiate second mesiobuccal (MB2) root canals in maxillary molars using nickel-titanium (NiTi) instruments in rotary and reciprocating kinematics. A total of 110 patients with MB2 canals were randomly assigned to two groups: EL - Easy Logic2 rotary instruments (n = 57) and X1B - MKLife X1 Blue reciprocating instruments (n = 53). Canals reached to full working length (FWL) were classified as "negotiable". In the X1B group, 96.2% of MB2 canals were negotiable, compared to 86% in the EL group, with no statistically significant difference (p = 0.152). Mean shaping time was shorter in the EL group (12.92 ± 4.36 min) than in the X1B group (14.33 ± 4.72 min), showing a significant difference (p = 0.047). Both systems showed high MB2 negotiability. However, regarding negotiability, both systems did not show statistically significant differences when adjusted Cox regression was used (Hazard ratio: 0.77; 95% confidence interval: 0.51 - 1.14). Rotary instruments showed a slight advantage in shaping time efficiency.
β-Thalassemia is a common hemoglobin disorder distributed worldwide and caused by mutations in the HBB gene that reduce or abolish β-globin chain synthesis, resulting in chronic hemolytic anemia, ineffective erythropoiesis, and variable clinical severity. Although the molecular spectrum of β-thalassemia has been extensively investigated, data correlating β-globin mutations with hematological characteristics in Iraqi patients remain limited. This study aimed to evaluate genotype-phenotype correlations by integrating molecular mutation analysis with hematological and biochemical profiling in an Iraqi β-thalassemia cohort. A cross-sectional study was conducted between September and November 2025 at the Hematology Department, University of Baghdad. Fifty patients with β-thalassemia were initially recruited, of whom 22 fulfilled the inclusion criteria and consented to participate (4 males and 18 females; age range, 18-68 years). Hematological parameters, HbA₂ and HbF levels, serum iron and ferritin concentrations, and β-globin gene mutations were analyzed. Mutation screening was performed using amplification refractory mutation system polymerase chain reaction (ARMS-PCR). Statistical analyses were conducted to evaluate genotype-phenotype associations. Patients exhibited a predominantly microcytic, hypochromic hematological profile, with a mean hemoglobin concentration of 11.1 ± 1.4 g/dL, a mean MCV of 63.5 ± 0.78 fL, a mean MCH of 20.2 ± 0.25 pg, and elevated RDW values (17.9 ± 0.23%), consistent with anisocytosis. HbA₂ levels ranged from 3.5% to 7.2% (mean, 5.4 ± 0.25%) as determined by HPLC, whereas HbF levels ranged from 0.3% to 5.6% (mean, 1.4 ± 0.17%). HPLC provided more reliable HbA₂ quantification than alkaline electrophoresis. Serum iron and ferritin levels showed considerable interindividual variability. β-Globin mutations were identified in 68.2% of patients, with IVS-I-6 being the most frequent mutation (45.5%), followed by CD39. No significant associations were observed between genotype and sex, profession, ethnicity, or place of residence. In contrast, age-group distribution differed significantly. This study demonstrated considerable phenotypic heterogeneity among Iraqi patients with β-thalassemia and identified IVS-I-6 as the predominant β-globin mutation in this cohort. The absence of significant genotype-phenotype associations for most clinical variables suggests that additional genetic and environmental modifiers may contribute to disease expression. Combined molecular and hematological evaluation may improve the diagnostic characterization and classification of patients with β-thalassemia.
Primary hyperparathyroidism is one of the most common endocrine diseases characterized by impaired calcium-phosphorus metabolism due to hypersecretion of parathyroid hormone. Assessment of the epidemiological situation with parathyroid gland diseases in the Chuvash Republic based on data from the All-Russian Registry of Patients with primary hyperparathyroidism. To analyze the data of patients with primary hyperparathyroidism in a retrospective cohort study from 2014 to 2025 at the clinical base of the Republican Clinical Hospital for War Veterans of the Ministry of Health of Chuvashia. The data on the Chuvash Republic from the All-Russian Registry of patients with primary hyperparathyroidism are analyzed. In the course of a retrospective cohort study (2014-2025), a cohort of patients registered at the Republican Clinical Hospital for War Veterans of the Ministry of Health of Chuvashia was analyzed. For statistical data processing, a descriptive method was used with the calculation of absolute and relative values (percentages). It was found that the prevalence of parathyroid gland pathology in the region is 73.4 cases per 100,000 population. Key epidemiological features were identified: a pronounced predominance of women (88.9%) in the older age group, an uneven geographical distribution with a concentration of cases in urbanized areas (Cheboksary, Novocheboksarsk), as well as a significant increase in detection in recent years. The data obtained indicate the influence of both organizational factors (availability of specialized care) and possible environmental risks (imbalance of trace elements in water sources) on the prevalence of pathology. The implemented electronic registry has proven its effectiveness for monitoring morbidity, planning medical care and conducting scientific research, which determines the prospects for its further development to optimize the system of endocrinological care in the region.
This study evaluated the proximate composition, fatty acid profile, mineral composition, and heavy metal contamination of brown bear (Ursus arctos) muscle, with emphasis on its nutritional quality and potential health risks for consumers. Muscle samples (musculus semimembranosus and musculus triceps brachii; n = 10) obtained from free-ranging individuals in Slovakia were analysed using validated analytical methods. Brown bear muscle was characterised by high protein content (~ 22.5 g/100 g) and low intramuscular fat content (< 1 g/100 g), confirming its lean nutritional profile. The fatty acid composition was dominated by monounsaturated fatty acids, followed by saturated and polyunsaturated fatty acids. Lipid quality indices indicated favourable nutritional properties, including low atherogenic and thrombogenic indices and a PUFA/SFA ratio above recommended nutritional thresholds. However, the relatively high n-6/n-3 ratio reflected the omnivorous feeding ecology and seasonal physiological status of the analysed animals. Iron and zinc represented the predominant trace minerals, highlighting the nutritional value of the analysed muscle tissue. Most toxic elements (As, Cr, Cd, Hg) remained below analytical detection or quantification thresholds and were therefore interpreted descriptively. Lead (Pb) was detected at low mean concentrations; however, substantial inter-individual variability resulted in sporadically elevated values, most likely associated with secondary contamination from bullet fragments. Exposure assessment based on consumption of a 100 g portion and an upper-bound approach for left-censored data indicated negligible toxicological relevance for Cd and Hg, whereas Pb exposure increased considerably under maximum exposure scenarios. Overall, brown bear muscle represents a nutritionally valuable food source; however, continued monitoring of Pb contamination in game meat remains warranted.
This study investigated the effects of selenium supplementation in the form of selenium nanoparticles (SeNPs) and sodium selenite (SSe) on the transcriptional regulation of heat shock proteins (HSP60, HSP70, and HSP90) in periparturient cloned and non-cloned Saanen goats and their offspring. A total of 24 pregnant goats (12 cloned via somatic cell nuclear transfer and 12 non-cloned) were divided into three groups: SeNPs, SSe and control. Supplementation was administered orally from 21 days before expected parturition. Serum selenium levels and HSP gene expression were measured at multiple time points relative to parturition. Both SeNPs and SSe significantly increased serum selenium levels, with SeNPs demonstrating superior bioavailability. Supplemented groups exhibited significantly lower transcriptional levels of HSP60, HSP70 and HSP90 during the periparturient period, with SeNPs showing a more potent suppressive effect. A significant negative correlation was observed between serum selenium and HSP transcription. No differences were found between cloned and non-cloned animals in selenium levels or HSP expression. These findings suggest that selenium supplementation, particularly as SeNPs, enhances antioxidant capacity and reduces cellular stress, as reflected by the downregulation of HSP genes. This study highlights the potential of nano-selenium to improve stress resilience during the critical transition period in goats and their offspring, irrespective of cloning status.
In Arabidopsis thaliana, BOR1 and BOR2 are well-characterized borate exporters. The role of the paralog BOR3, however, has remained undetermined. We found that the bor1 bor2 bor3 triple mutant exhibits significantly poorer root elongation than the double bor1 bor2 mutant, suggesting that BOR3 contributes to the uptake of boron from the soil to the inner tissues in the absence of BOR1 and BOR2. In contrast to BOR1 and BOR2, BOR3 localization in the plasma membrane of the root cells was not noticeably polar, as would be expected given its ability to promote directional boron transport under the triple mutant background. To understand this apparent dichotomy between function and localization, we modeled the effect of BOR3 polarity on boron flow. It revealed that a synergy of diffusive processes can lead to directional transport within a plant tissue. The counterintuitive mechanism, here termed diffusive alliance transport (DAT) consists of perfectly apolar (or even outward-facing) exporters working in conjunction with polarly localized diffusion facilitators to enable the buildup of higher-than-medium concentrations as well as inward-directional flux within the tissue. For this mechanism to be operational, the existence of apoplastic compartments between cells is essential.
Avian pathogenic Escherichia coli (APEC) causes significant economic losses in poultry production, yet the molecular mechanisms underlying divergent infection outcomes remain poorly defined. This study aimed to elucidate the in vivo transcriptional profiles in APEC from infected chicken embryos compared to in vitro grown culture. Chicken embryos were experimentally infected with an APEC isolate. Subsequently, RNA sequencing was performed on APEC isolated from the yolk of experimentally infected embryos that either died or survived the infection, alongside in vitro grown controls. Differential gene expressions, functional enrichment analyses, small RNA profiling were conducted to identify key transcriptional changes. Weighted gene co-expression network analysis (WGCNA) was applied to detect bacterial gene modules associated with host outcomes. In embryos that died, APEC exhibited a transcriptionally inferred hypermetabolic program, characterized by upregulation of 443 genes involved in anaerobic respiration, nutrient scavenging, and purine and arginine biosynthesis, consistent with increased replication and virulence potential. In contrast, APEC recovered from surviving embryos displayed a transcriptionally inferred low-energy state under host immune pressure, characterized by suppression of central metabolism pathways and concurrent upregulation of genes associated with iron sequestration, toxin production, and cell envelope remodeling. WGCNA suggested the association of low-oxygen respiration and anaerobic metabolism, Fe-S cluster assembly, and purine and arginine biosynthesis genes with host lethality, while siderophore-mediated iron acquisition correlated with host survival. Arginine metabolism showed a positive association with pro-inflammatory host responses, while flagellar motility was associated with both pro- and anti-inflammatory signatures, suggesting a modulatory role of metabolic and motility pathways on host immunity. The analyses indicated that APEC survival and pathogenicity are more dependent on metabolic adaptation and iron competition than individual toxin genes. Additionally, 77 small regulatory RNAs were identified, suggesting potential regulatory roles in in vivo adaptation.These findings demonstrate the transcriptional plasticity of APEC during infection, highlighting that low-oxygen respiration and anaerobic metabolism, iron acquisition, arginine and purine metabolism and flagellar motility may contribute to bacterial survival strategies. These findings are consistent with recent host transcriptomic evidence showing mitochondrial dysfunction, hyperinflammation, and metabolic collapse in the yolk sac of infected embryos, suggesting that coordinated host-pathogen metabolic interactions influenced infection outcomes.
Heavy metal contamination of agricultural soils severely impairs plant physiological function by disrupting nutrient uptake, photosynthesis, and redox homeostasis. Previous studies have established that heavy metals such as cadmium (Cd), chromium (Cr), lead (Pb), and copper (Cu) induce oxidative stress, membrane damage, and metabolic imbalance in plants, and that flavonoids can act as antioxidants to mitigate some of these effects. However, the specific mechanistic role of hesperidin in regulating plant physiological responses under heavy metal stress in canola remains unclear. In this study, we investigated the effects of exogenous hesperidin application on canola (Brassica napus L.) exposed to Cd, Cr, Pb, and Cu stress. Hesperidin treatment enhanced enzymatic antioxidant systems, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), as well as non-enzymatic antioxidants, thereby reducing reactive oxygen species (ROS) accumulation and stabilizing membrane integrity. In addition, hesperidin regulated osmolyte metabolism (proline and soluble sugars) and restored ion homeostasis, leading to improved nutrient acquisition and photosynthetic efficiency. These coordinated responses demonstrate that hesperidin functions as a physiological regulator, integrating antioxidant defense and metabolic adjustment to confer resilience against heavy metal toxicity. The novelty of this work lies in identifying hesperidin's dual role in modulating both redox balance and osmolyte metabolism, thereby providing new mechanistic insight into flavonoid-mediated stress tolerance. Our findings highlight hesperidin as a potential biotechnological tool to improve crop performance and resilience in contaminated environments.
Photon-counting CT (PCCT) enables K-edge imaging, an advanced multi-material decomposition technique that requires multiple energy bins to perform decomposition unconstrained by physical assumptions. The clinical implementation of K-edge imaging applications like dual-contrast imaging could lead to reductions in radiation dose and motion artifacts between sequential scans. To evaluate the feasibility and performance of K-edge imaging of iodine (I) and gadolinium (Gd) on a clinically available photon-counting computed tomography (PCCT) system. A dual-source clinical PCCT scanner with four energy thresholds (20, 55, 72, 90 keV) was used to scan phantoms containing single and dual contrast solutions of I and Gd across multiple concentrations (1-10 mg/mL) and radiation doses (1-8 mGy). Multi-material decomposition was performed using a calibration-based, image-domain algorithm to generate material-specific maps. Quantitative accuracy was assessed using Bland-Altman analysis and contrast-to-noise ratio (CNR), while noise and bias trends were statistically analyzed using non-parametric tests. K-edge imaging was successfully achieved on a clinical PCCT system with accurate decomposition of I and Gd across varying concentrations, solution compositions (single/dual contrast), and dose levels. Quantitative bias was significantly influenced by radiation dose, concentration, and solution composition (p < 0.0004). Increased radiation dose and contrast concentration improved quantification accuracy, with maximum bias reductions of 0.9 (I) and 0.3 mg/mL (Gd). CNR correlated linearly with concentration (R2 > 0.99) and moderately with dose (R2 = 0.85-0.94), achieving peak values of 13 (I) and 16 (Gd) at 8 mGy. Dual contrast solutions showed reduced performance compared to single contrast solutions, that is CNR of 5 mg/mL Gd solutions increased by 0.6 per mGy in single contrast solutions while by 0.5 per mGy in dual contrast mixtures. Noise was dependent on dose but not on concentration or solution composition. This study establishes the feasibility of K-edge imaging using a clinical PCCT system and demonstrates accurate, simultaneous decomposition of I and Gd across composition of solution. These findings support the clinical translation of K-edge imaging and highlight PCCT's potential for advanced dual-contrast and molecular imaging applications.
Cobalt stress can promote the accumulation of dendrobine-type total alkaloids (DTTAs) in Trichoderma longibrachiatum UN32; however, the signalling mechanisms underlying this response remain unclear. In this study, we examined whether changes in Ca2+ and nitric oxide (NO) signalling were associated with ROS-related responses and cobalt-enhanced DTTAs production under cobalt stress. Co2+ treatment increased DTTAs accumulation together with intracellular hydrogen peroxide (H2O2), Ca2+ and NO levels. Among Ca2+ signalling inhibitors, neomycin (Neo) showed the strongest effect: 6 mM Neo reduced intracellular Ca2+ by 59.86%, DTTAs production by 33.70% and H2O2 levels by 25.50% compared with CoCl2 treatment. Time-course analysis further showed coordinated changes in H2O2, Ca2+ and DTTAs accumulation during fermentation. In contrast, L-NAME-mediated inhibition of NO synthesis did not significantly affect DTTAs production. These findings suggest that ROS-Ca2+ interplay is associated with cobalt-enhanced DTTAs accumulation in T. longibrachiatum UN32, whereas NO has a limited role under the tested conditions.
Chemoselective bioconjugation techniques are essential tools for precise modification and functional analysis of proteins. Here, we describe a detailed protocol for histidine-specific labeling using thiophosphorodichloridate reagents, forming stable thiophosphoramidate linkages. The procedure includes synthesis of the labeling reagents, protein modification, purification by ion-exchange chromatography, and subsequent copper-catalyzed azide-alkyne cycloaddition (CuAAC) for further conjugation.