Infections caused by multidrug-resistant Gram-negative bacteria, particularly carbapenemase-producing Enterobacterales, present significant diagnostic and therapeutic challenges, highlighting the need for practical laboratory methods to evaluate combination therapies. This study investigated 90 ceftazidime-avibactam (CZA)-resistant Klebsiella pneumoniae isolates collected from clinical specimens between January 2021 and July 2023. Species identification was performed using MALDI-TOF MS, and antimicrobial susceptibility testing was conducted with the BD Phoenix™ system according to Clinical and Laboratory Standards Institute (CLSI) criteria, with disk diffusion used for confirmation when necessary. Carbapenemase production was determined phenotypically using the modified Carbapenem Inactivation Method. For a subset of isolates with available whole-genome sequencing data, resistance genes were confirmed, although genotypic analysis was not performed for all isolates. The in vitro activity of the CZA plus aztreonam (CZA-ATM) combination was evaluated using the CLSI-recommended broth disk elution method. All isolates were resistant to CZA and aztreonam when tested individually, whereas no visible growth was observed with the combination, indicating in vitro susceptibility. These findings suggest that the broth disk elution method is a practical approach for assessing CZA-ATM activity in routine clinical laboratories. Further studies including reference method validation and evaluation of borderline or resistant isolates are needed.
In this study, a methodology has been developed for the estimation of the spatiotemporal distribution of 137Cs dose rates in four widely consumed pelagic and pelagic-neritic fish species, Boops boops (Gopa), Spicara flexuosum (Tseroula), Sardina pilchardus (Sardella), and Trachurus trachurus (Savridi), in the South Aegean Sea, Greece, based on earth observation (EO) data. These species are selected due to their native status in the Mediterranean, abundance in the Aegean Sea, and their representation of different habitat types within pelagic and pelagic-neritic systems. The 137Cs surface seawater activity concentrations are estimated using previously established equations derived from relations between EO marine parameters and laboratory measurements of 137Cs seawater activity concentrations. These EO marine measurements, namely NASA's MODIS iPAR and PAR (photosynthetically available radiation) level 3 monthly products, are used as input in these equations for the estimation of monthly surface seawater 137Cs activity concentration maps, within a GIS environment. The corresponding dose rate maps for each one of the four fish species are produced by using well-established radiation protection equations and the estimated 137Cs surface seawater activity concentration maps: (a) assuming external dose rate in an infinite space and (b) internal dose by using species-specific concentration factors (CFs), which have been calculated for the Aegean Sea in ENRACT, enabling assessment of variability in radionuclide uptake and associated dose rates. The validity of these estimations is evaluated through independent 137Cs activity concentration measurements from the South Aegean Sea, while uncertainty was assessed performing a sensitivity analysis. The approach is limited by the region-specific applicability of the regression model and the lack of sufficient validation datasets. The produced monthly maps cover the years 2017-2019 with a spatial resolution of 4 km. The results demonstrate the potential of integrating EO data with radioecological modeling in marine ecosystems for producing routine monitoring maps on a systematic time scale.
Bone sarcoma treatment employs wide surgical margins to ensure complete tumour excision, sometimes at the expense of patient quality of life. As precision technologies advance, quantitative data to inform the planned accuracy of surgical margins is increasingly important. This study assesses two novel computational methods (M2, M3) to support the co-registration of 2D histologic images within 3D radiological volumes. Replicated evaluation of an established method (M1) is also performed with new data. M2 suits concave bisected sections by assuming a different orientation during tissue bandsaw than M1. M3 handles freeform geometries by using a third measurement to constrain 2D histology images within 3D space without need for geometric bandsaw assumptions. Three computed distances across 38 canine specimen dissections from each method were compared with three corresponding physical measurements collected during laboratory processing. M1 produced a mean error of 0.12 (± 1.7) mm, M2 produced a mean error of 0.06 (± 1.9) mm, while M3 produced a mean error of 0.02 (± 1.5) mm. Mean errors for each method were of similar magnitude to 3D registration error reported in histology co-registration frameworks for other tissues. This study supports a simple low cost opportunity for use of digital histology images to support bone sarcoma radiology interpretation and treatment planning.
Intradialytic hypotension (IDH) is a frequent complication of maintenance hemodialysis and is associated with increased morbidity. The role of adrenal insufficiency (AI) as a potential contributor to recurrent IDH remains insufficiently characterized. To evaluate the prevalence of adrenal insufficiency among hemodialysis patients with recurrent intradialytic hypotension and to examine associated clinical and biochemical characteristics. In this cross-sectional study, adult patients undergoing maintenance hemodialysis who experienced recurrent intradialytic hypotension were recruited from a tertiary dialysis center. Morning serum cortisol and plasma adrenocorticotropic hormone (ACTH) were measured prior to dialysis. Patients were classified as having suspected adrenal insufficiency if their serum cortisol levels were less than 18 μg/dL. Clinical and laboratory parameters were compared between adrenal insufficient and adrenal-sufficient patients. Fifty-one hemodialysis patients with recurrent intradialytic hypotension were evaluated. The mean age was 52.35 ± 13.6 years, and 37.2% were male. Adrenal insufficiency was identified in 37 patients (72.5%). Based on basal ACTH levels, 4 patients (7.8%) were classified as suspected primary AI and 33 (64.7%) as suspected secondary AI. No significant differences were observed between groups regarding age, sex, dialysis vintage, or interdialytic weight gain. Adrenal insufficient patients demonstrated significantly lower serum sodium and random blood glucose levels and higher serum potassium levels compared with adrenal-sufficient patients (p < 0.05). Adrenal insufficiency was frequently detected among hemodialysis patients with recurrent intradialytic hypotension in this single-center cohort. Biochemical findings were compatible with impaired adrenal function. Further studies are required to validate diagnostic thresholds in this population and to determine the therapeutic implications of identifying adrenal dysfunction in patients with intradialytic hypotension.
Semantic-categorically related distractor words slow down picture naming more than semantically unrelated distractor words do (e.g., the words "dog" vs. "car" for a picture of a horse). This semantic interference effect in the picture-word interference task has been replicated numerous times and is an empirical cornerstone of speech production research. However, studies using the picture-picture interference task, in which the distractor words are replaced with distractor pictures, have yielded highly inconsistent results, reporting semantic interference, semantic facilitation, and null effects. Here, we present a meta-analytic summary of the results of published picture-picture interference studies. We found the estimated semantic effect to be close to zero (2 ms, 95% confidence interval [CI] [-5, 9]). This contrasts with the results of 14 previously unpublished picture-picture interference experiments conducted in our laboratory and which we report here. Across these experiments, the estimated semantic-interference effect was 15 ms (95% CI [9, 21]). When these data were included in the meta-analysis, the pooled estimate was 9 ms (95% CI [4, 14]). These results suggest that the magnitude of semantic interference from distractor pictures has been underestimated and that it is not as elusive as previously thought. We discuss the implications for modeling word production. (PsycInfo Database Record (c) 2026 APA, all rights reserved).
Objective laboratory diagnostics for autism spectrum disorder (ASD) are lacking, necessitating rapid clinical screening tools. Because serum extracellular vesicle (EV) N-glycosylation captures critical neurodevelopmental signatures, we developed a fast, biologically interpretable diagnostic strategy. EVs from ASD patients with language impairment and neurotypical controls were isolated using a rapid extra-polyethylene glycol precipitation/filtration (EPF) workflow, benchmarked against ultracentrifugation. Following MALDI-TOF/MS profiling, machine learning was re-evaluated using repeated nested cross-validation to reduce optimistic bias and potential information leakage. Among five classifiers, Random Forest (RF) showed the best overall balance across discrimination, calibration, and classification metrics. RF-based SHAP analysis provided transparent interpretation, highlighting key discriminative glycans, including H4N3S1F1, H5N5S1F1, and H3N5F1. To elucidate molecular mechanisms, we integrated public EV transcriptomic data. This revealed significant dysregulation of N-glycosylation machinery genes (e.g., MAN1A1, NEU1, OSTC, RPN2), whose expression directionally aligned with observed glycan shifts in synaptic pathways. Collectively, this rapid serum EV N-glycomic workflow, combined with leakage-controlled RF-based interpretation, provides a promising foundation for non-invasive ASD biomarker discovery and future multicenter validation.
After the COVID-19 emergency phase, Mycoplasma pneumoniae (MP) infections surged globally, heightening concerns over macrolide-resistant M. pneumoniae (MRMP), especially in children. Early detection of MRMP is crucial for reducing morbidity and optimizing antimicrobial use, but developing reliable prediction models remains challenging. This retrospective study evaluated 458 hospitalized children with PCR-confirmed MP pneumonia during the 2024 outbreak at a South Korean university hospital. We compared demographic, clinical, laboratory, radiographic, and treatment data between macrolide-sensitive (MS) and -resistant groups. Multivariable logistic regression and ROC analyses identified independent predictors of resistance and developed a risk model. The prevalence of MRMP was profoundly high at 83.8%. Compared with MS infections, MRMP was associated with older age, higher inflammatory markers, lower MP PCR cycle threshold (Ct) values, and more pulmonary complications. Multivariable analysis identified elevated erythrocyte sedimentation rate (ESR) and lower PCR Ct as robust, independent predictors of macrolide resistance. Integrating the PCR Ct value into a baseline clinical model significantly improved prediction accuracy, increasing the area under the curve (AUC) from 0.659 to 0.806. Chest radiographic findings provided negligible incremental diagnostic value (AUC 0.808). MRMP is highly prevalent among hospitalized children during post-emergency outbreaks in East Asia. Incorporating admission PCR Ct values and ESR into early clinical assessments substantially improves prediction of macrolide resistance, enabling timely, targeted escalation to second-line therapies.
Current recommendations for screening Chlamydia trachomatis (CT) and Neisseria gonorrhoeae in urinary tract infections using nucleic acid amplification tests (NAATs) include vaginal swabs for women and first‑catch urine for men. In addition, some manufacturers require the use of a transport medium for prolonged pre‑analytical storage. The aim of this study was to evaluate the sensitivity and specificity of midstream urine specimens and to assess nucleic acid stability in neat urine samples stored at +4°C. The study included 338 asymptomatic participants undergoing screening for Chlamydia trachomatis and Neisseria gonorrhoeae as part of the French national STI screening program (172 women and 166 men). For each participant, a reference specimen (self‑collected vaginal swab for women and first‑catch urine for men) was compared with a midstream urine sample. Two comparisons were conducted to assess nucleic acid stability: the first compared urine stored in a preservation medium with neat urine stored at +4°C, and the second compared neat urine stored at +4°C on days 1 and 7. All samples were analysed using the Alinity m Abbott® automated system with the STI AMP Kit. Among the 46 positive samples, the prevalence of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) was 43/338 and 4/338, respectively. Due to the low prevalence of NG, sensitivity and specificity analyses for midstream urine were limited to CT. Vaginal swabs showed superior performance for detecting CT in females, with two infections missed by midstream urine. Midstream urine demonstrated adequate sensitivity in females (92.9%) and excellent sensitivity in males (100%), with no false-positive results observed. Regarding nucleic acid stability, no statistical difference (p=0.799) was observed for cycle values between urine samples eluted into the liquid medium and neat urine stored at +4°C. Furthermore, our study validated the use of prolonged storage periods (up to 7 days) at +4°C for CT and NG detection in neat urine (p=0.671). In conclusion, the results of this study corroborate the CDC's 2014 recommendations, but the use of midstream urine could offer interesting prospects for Chlamydia trachomatis screening in specific populations. Regarding Neisseria gonorrhoeae, given the low prevalence of positive results found in our study, no meaningful conclusions can be drawn.
Gastric cancer (GC) is one of the leading causes of cancer-related deaths, particularly in early-stage GC, which is frequently asymptomatic, and there are no reliable molecular markers available for diagnosis. To identify potential candidate genomic biomarkers for GC with respect to genetic variations and expression changes in genes related to hydrogen sulfide (H2S) metabolism and antioxidant defence. We included 15 patients with newly diagnosed gastric adenocarcinoma in a paired case-control molecular design based on tumour tissue and adjacent histologically normal gastric mucosa. Biopsy tissues were used to extract genomic DNA and total RNA. Complementary DNA (cDNA) was synthesised, followed by standard polymerase chain reaction, Sanger sequencing, and quantitative real-time polymerase chain reaction analysis. The selected target genes were cystathionine beta-synthase (CBS), cystathionine gamma-lyase (CTH), 3-mercaptopyruvate sulfurtransferase (MPST), catalase (CAT), and glutathione peroxidase 1 (GPX1). A total of 53 insertion/deletion/duplication variants without amino acid change and 11 predicted amino acid-changing sites plus multiple homozygous substitution variants were identified through sequence variants screening. Most variants were heterozygous and not present in external databases. Common high-frequency variants included CBS 19,430 C > CG, CTH 28400G > GT, CAT 33470G > GA, and changes associated with GPX1613_618delGGCGGC. Expression analysis indicated upregulation of CBS, CAT, and GPX1 (by approximately 1.40-, 1.73-, and 1.93-fold, respectively), downregulation of MPST (3.11-fold), and slight downregulation of CTH. Somatic mutations and altered expression of CBS, CTH, MPST, CAT, and GPX1 in GC tissues indicate coordinated remodeling of sulfur metabolism and redox-defence pathways. Nevertheless, the low ROC performance suggests that these genes should not yet be viewed as viable solo diagnostic biomarkers. Instead, they are exploratory molecular signatures and candidate markers that must be validated on larger series of independent patients, matched tumour-normal sequencing, protein/enzyme lateral re-confirmation and functional studies.
In this study, the transcriptional coactivator ZaGIF2, isolated from Zanthoxylum armatum, enhances plant growth and drought tolerance by synergistically activating GRF transcription factors in plants. Growth-regulating factor-interacting factors (GIFs) are conserved transcriptional coactivators in plants that regulate growth and development by forming complexes with growth-regulating factors (GRFs). However, their functions in woody plants, especially in non-model species, and their specific roles in coordinating growth and stress resistance remain unclear. In this study, a GIF gene, ZaGIF2, was cloned from the economically important species Zanthoxylum armatum var. dintanensis. ZaGIF2 exhibited the highest expression in apical buds with active meristems, and its transcription was significantly induced by naphthaleneacetic acid (NAA), abscisic acid (ABA), and drought stress. The encoded protein was localized to the nucleus. Overexpression of ZaGIF2 in Nicotiana benthamiana and Arabidopsis thaliana simultaneously promoted leaf enlargement and enhanced drought tolerance. Transgenic plants demonstrated improved water retention capacity, reduced oxidative damage, and enhanced osmotic adjustment. Overexpression of ZaGIF2 upregulated the expression of multiple growth-related genes (AtPIF4, AtLBD18, AtTCP14) and drought-responsive genes (DREB2A, P5CS1, RD26, RD29). Mechanistically, the ZaGIF2 protein directly interacted with AtGRF1 and AtGRF3 and significantly enhanced their transcriptional activation activity. Collectively, our findings reveal a novel mechanism in woody plants whereby a GIF protein improves plant growth and drought adaptation by interacting with and enhancing the activity of GRF transcription factors. This study provides important genetic resources and a theoretical foundation for the synergistic improvement of growth and stress resistance traits in forest trees using the conserved GIF-GRF module.
Glucagon is a key regulator of amino acid metabolism, and elevated levels are frequently observed in maintenance hemodialysis (MHD) patients. Yet, the relationship between plasma glucagon levels, nutritional parameters, and prognosis in MHD patients remains unclear. We evaluated whether glucagon levels in MHD patients are associated with normalized protein catabolic rate (nPCR), glucagon-like peptide-1 (GLP-1), nutritional status, and all-cause mortality. In this prospective observational study, plasma hormone levels were measured alongside clinical, biochemical, anthropometric, bioimpedance-derived nutritional markers, and inflammatory indices. Mortality was assessed using Kaplan-Meier and Cox regression analyses. This study included 82 prevalent MHD patients. Patients were followed for 18.0 ± 4.9 months. Glucagon correlated positively with nPCR (r = 0.25, p = 0.03) and inversely with phase angle (PhA) (r =  - 0.24, p = 0.03) after adjustment. Additional correlations with GLP-1 became marginal after adjustment. The subgroup with high glucagon and low nPCR showed the lowest PhA (p = 0.03), and remained significant after multivariable adjustment. Patients with glucagon ≥ 120 pg/mL (ROC-derived cutoff) had higher mortality (log-rank p = 0.05); however, the discriminative power of this cutoff was limited (AUC 0.63). In multivariable models, the association persisted after adjustment for age and diabetes but was attenuated after further controlling for PhA, GLP-1, or MIS. Elevated glucagon in MHD patients was modestly associated with altered protein metabolism, impaired nutritional integrity, and borderline higher all-cause mortality. The independent prognostic value of glucagon remains uncertain, and a concept of a glucagon resistance phenotype is speculative.
State transitions balance excitation energy between photosystem II and photosystem I and represent a key photoprotective mechanism in oxygenic photosynthesis. While extensively characterized in model cyanobacteria, the regulation of state transitions in diazotrophic species remains poorly understood. Here, we present a comparative physiological study of two unicellular nitrogen-fixing cyanobacteria, Crocosphaera watsonii WH 8501 and Crocosphaera subtropica ATCC 51142 under diazotrophic and non-diazotrophic conditions. Using growth analyses, whole-cell absorption spectroscopy, and low-temperature (77 K) fluorescence spectroscopy, we examined how nitrogen availability shapes photosynthetic energy distribution across species and conditions. Our results showed that state transitions are tightly linked to nitrogen metabolism in both species, but with distinct species-specific regulatory strategies. Diazotrophic cultures exhibited a higher capacity for state transitions than non-diazotrophic cultures, indicating enhanced flexibility in excitation energy redistribution. In C. watsonii, pronounced state transitions were observed exclusively under diazotrophic conditions, whereas non-diazotrophic cultures relied on alternative modes of excitation regulation. In contrast, C. subtropica displayed a more flexible response, with state transitions influenced by both nitrogen availability and light intensity. Fluorescence analyses revealed that state transitions involve a combination of excitation spillover between photosystems and dynamic coupling-decoupling of phycobilisomes. Together, these findings suggest that nitrogen fixation is associated with changes in photosynthetic energy regulation in marine diazotrophic cyanobacteria.
Developing radiosensitizing agents to amplify tumor-eradicating effects on primary, regional recurrence, and distant metastases plays a transformative role in modern cancer care. Here, we report the de novo design of biocatalytic artificial metalloenzymes with an IrMn-cluster-based redox center (IMM) to achieve radiosensitized systemic antitumor responses for preventing malignant tumor metastasis and recurrence. Notably, our findings indicate that Mn-organic ligands substantially enrich the electron density of Ir clusters, thereby optimizing their interaction with oxygen species and markedly enhancing the production of both reactive oxygen species and molecular oxygen. When combined with radiotherapy, the IMM effectively amplifies DNA damage and induces pronounced apoptosis by alleviating intratumoral hypoxia. This shift reprograms the tumor microenvironment, enhancing radiosensitivity and facilitating the infiltration and activation of intratumoral CD8⁺ T cells and dendritic cells. Moreover, when integrated with anti-PD-1 therapy, this coordinated therapeutic regimen elicits potent systemic immune responses and durable antitumor memory, effectively suppressing tumor recurrence and metastasis while markedly improving therapeutic efficacy and long-term survival. We anticipate that this conceptual design could offer a promising and translationally relevant nanomedicine platform for radiotherapies.
The optimal strategy for laryngeal preservation in locally advanced hypopharyngeal carcinoma (LAHPC) remains undefined. This prospective observational study compared long-term outcomes between two primary treatment approaches: induction chemotherapy followed by concurrent chemoradiotherapy (IC-CCRT) and neoadjuvant chemotherapy followed by surgery and adjuvant (chemo)radiotherapy (NAC-Sur-RT). From December 2018 to December 2023, 199 patients with LAHPC were prospectively enrolled and treated with either IC-CCRT (n = 124) or NAC-Sur-RT (n = 75). The primary endpoints were survival with functional larynx (SFL), overall survival (OS), progression-free survival (PFS), locoregional failure-free survival (LRFFS), and distant metastasis-free survival (DMFS). Prognostic factors were identified using multivariable Cox regression. In the matched cohort (median follow-up 49 months), NAC-Sur-RT was associated with significantly superior 5-year OS (58.3% vs 46.2%; adjusted HR = 0.60; 95% CI 0.38-0.95; P = 0.031) and DMFS (56.2% vs 41.6%; adjusted HR = 0.59; 95% CI 0.38-0.92; P = 0.023), and PFS (52.2% vs 37.2%; adjusted HR = 0.64; 95% CI 0.42-0.98; P = 0.040). Conversely, NAC-Sur-RT was associated with a significantly higher risk of SFL failure (27.0% vs 43.1%; adjusted HR = 2.18; 95% CI 1.45-3.27; P < 0.001). No significant differences were found in LFFS or LRFFS. Failure pattern analysis indicated more isolated local recurrences with IC-CCRT (15.3% vs 5.3%) but higher regional recurrence with NAC-Sur-RT (1.6% vs 6.7%). Multivariable analysis confirmed NAC-Sur-RT as an independent predictor of improved OS (HR = 0.60; P = 0.031), PFS (HR = 0.64; P = 0.040), and DMFS (HR = 0.59; P = 0.023), but also of worse SFL (HR = 2.18; P < 0.001). AJCC stage IVB disease independently predicted inferior OS (HR = 9.05; 95% CI 1.52-53.81; P = 0.015) and DMFS (HR = 8.49; 95% CI 1.53-47.23; P = 0.015). Additionally, NAC-Sur-RT was associated with significantly higher rates of severe (grade 3-4) toxicities, including laryngeal/esophageal, mucosal, hematologic, and functional adverse events. In this non-randomized prospective comparison, the NAC-Sur-RT strategy was associated with superior survival outcomes (OS, PFS, DMFS) but at the cost of increased severe toxicity and a significantly lower rate of functional larynx preservation. IC-CCRT offers a higher probability of survival with a functional larynx despite a higher risk of local recurrence. Treatment selection should be individualized, balancing survival benefits, functional outcomes, and toxicity profiles. The study was approved by the ethics board of the Chinese PLA General Hospital and registered in the Chinese Clinical Trial Registry (registered number: ChiCTR2000039813).
Chalcopyrite is the most abundant copper-bearing sulfide mineral, characterized by a stable crystal structure. Bioleaching of chalcopyrite is inherently slow and further constrained by surface passivation layers during prolonged bioleaching. In this study, bacteriophages released from Acidithiobacillus ferrooxidans were introduced to promote chalcopyrite bioleaching. Mineralogical characterization, microbial profiling, and functional gene analysis were employed to reveal the underlying mechanisms. One day after phage introduction, phage abundance was 9.42 × 105 VLPs mL-1 in the phage-treated group, 150-fold higher than in the control (6.26 × 103 VLPs mL-1). Over a 60-day leaching period, the copper leaching efficiency of the phage-treated group was 31.72%, significantly higher than that of the control (20.43%). Phage introduction alleviated surface passivation, suppressed elemental sulfur (S0) and jarosite-type deposition, and increased the apparent product-layer diffusion rate constant 3.60-fold. Bacterial diversity and the relative abundance of heterotrophic bacteria were elevated. The increase in heterotrophic bacteria may alleviate the inhibitory effects of organic matter on autotrophic bioleaching microorganisms. Genes associated with the Sox system, dissimilatory sulfur metabolism, and organic sulfur transformation were significantly enriched in the phage-treated group, consistent with enhanced S0 turnover and reduced surface accumulation. Together, these results indicate that phage introduction enhances chalcopyrite bioleaching by alleviating surface passivation and promoting sulfur turnover through coupled interfacial and microbial regulation. This study provides a novel approach for sustainable intensification of bioleaching of refractory sulfide minerals.
Highly polar contaminants are analytically challenging due to the substantial differences in their chromatographic retention and ionisation behaviour between compound groups. This communication evaluates a developed and validated ZICcHILIC-HRMS workflow as a complementary approach to an established anion IC-HRMS method for analysing selected highly polar contaminants in water. The study focuses on implementing polarity switching, comparing method-specific analyte coverage and deriving practical method-selection criteria. Polarity switching enabled the acquisition of both positively and negatively ionising compounds within a single ZICcHILIC-HRMS run, thereby improving the practical efficiency of the comparatively lengthy chromatographic workflow. A comparison with IC-HRMS revealed partially overlapping, yet distinct, analytical coverage. IC-HRMS remained advantageous for small, strongly polar anionic compounds due to direct injection, lower quantification limits, shorter runtime and higher routine robustness. In contrast, ZICcHILIC-HRMS extends the accessible analyte spectrum to compounds outside the scope of the applied anion IC-HRMS and to analytes that can be detected in positive electrospray ionisation mode. These findings support the use of ZICcHILIC-HRMS as a complementary, LC-HRMS-compatible tool for the exploratory monitoring of highly polar contaminants in water, particularly when both ionisation modes and conventional HPLC-HRMS instrumentation are to be employed.
To determine the contrast enhancement values in the facial nerve segments using magnetic resonance imaging (MRI) and to examine the association with clinical progress in Bell's palsy. Temporal MRI images of 90 adult patients with Bell's palsy and 45 individuals without facial nerve pathology (control group) were analyzed retrospectively. Each of the five intratemporal segments of the facial nerve was delineated bilaterally, and quantitative values obtained using point-shaped ROI (region of interest) were recorded. These values were compared between the affected and unaffected sides of Bell's palsy patients, as well as between the unaffected side and the control group. The association between the disease's progress was examined. The ROI value of the affected side of the patients was significantly greater than the normal side in all segments. Contrast measures in five separate patient segments could not predict improvement after three months. At the end of the 6th month, ROI values could not predict whether the patient would have progression or not. In all 5 segments, ROI intensity values were found to be significantly higher in the unaffected sides of patients than in the ears of healthy individuals in the control group. The mean ROI intensity value in any segment of the facial nerve on the affected side could not predict the prognosis of patients with Bell's palsy. The considerable rise in signal intensity values evaluated by point-shaped ROI may suggest a possible role for systemic and local immunological responses; however, this interpretation remains a hypothesis, as no direct laboratory immunological evidence was obtained in the present study.
Stress contributes to disease outcomes and can influence the signaling and interactions of immune cells. This study aimed to evaluate the influence of three selected stress hormones cortisol, epinephrine and norepinephrine, at in vivo-relevant concentrations, on the antimicrobial activity of porcine neutrophils during infection with Actinobacillus pleuropneumoniae (A.pp). We analyzed whether stress hormones affect the antimicrobial activity of neutrophils. Reactive oxygen species (ROS) production, cell size, and granularity were measured using flow cytometry. Neutrophil extracellular trap (NET) formation was quantified using confocal immunofluorescence microscopy, and transmission electron microscopy was employed to analyze the formation of vesicular NETs. ROS production and NET formation experiments were also conducted under infection with A.pp. The survival of A.pp in the presence of stress hormone-treated neutrophils was assessed. The antimicrobial activity of neutrophils was altered in a concentration-dependent manner by all tested stress hormones: neutrophils produced higher amounts of ROS and released significantly more NETs under A.pp infection, in particular vesicular NET formation. However, A.pp growth in the presence or absence of neutrophils was not affected by stress hormones. Future studies are needed to characterize the role of vesicular NETs as stress response reactions. In conclusion, this study contributes to a better understanding of the innate immune cells, particularly porcine neutrophils, in infections within stressed hosts.
Salt stress induces ABI5 → ALKBH10B transcription; the demethylase removes m6A from RAP2.6 mRNA, accelerating its decay and attenuating salt-responsive genes, thus linking ABA signaling to reversible m6A control of Arabidopsis salt tolerance. N6-methyladenosine (m6A) is the most prevalent internal modification of RNA and plays an important role in regulating RNA metabolism that governs development and environmental adaptation of plants. Here we dissect how the Arabidopsis m6A demethylase ALKBH10B is integrated into abscisic-acid (ABA)-mediated salt-stress signaling. Under salt treatment, loss of ALKBH10B exhibited significantly delayed seed germination. Salt stress could significantly induce the expression of ALKBH10B transcription via the ABA-responsive transcription factor ABI5, which binds directly bind to the ABRE element in the ALKBH10B promoter and activate its transcription. Multi-omic integration of m6A methylomes and transcriptome identified the AP2/ERF transcription factor RAP2.6 as a direct target of ALKBH10B. ALKBH10B removed m6A modifications on RAP2.6 mRNA, accelerating its degradation and modulating the expression of salt-responsive genes. In summary, this study elucidates the salt stress response pathway ABI5-ALKBH10B-RAP2.6 that couples ABA perception to reversible m6A modification, providing mechanistic insight into the intricate regulatory network of dynamic m6A modifications in plant stress adaptation.
To explore the agreement among tear break-up time measurements performed by novice examiners using manual non-invasive tear break-up time (mNIBUT), automated non-invasive tear break-up time (aNIBUT) and fluorescein tear break-up time (FBUT), and to examine the association between tear film metrics and symptoms. A total of 61 volunteers (44 females) with a mean age of 21.5 ± 2.7 years underwent tear-film assessment performed by novice examiners (undergraduate students). The tear volume (Schirmer test), tear film stability by mNIBUT, aNIBUT and FBUT, along with lipid layer pattern, were assessed. Symptoms associated with tear-film quality and ocular surface discomfort were evaluated using the Ocular Surface Disease Index (OSDI) questionnaire. Normal values were seen in 88.5%, 93.4%, 73.8% and 39.3% of participants using the Schirmer test, FBUT, mNIBUT and aNIBUT, respectively. The intraclass correlation coefficient (ICC) for tear break-up measurements indicated poor reliability between tests, except when comparing mNIBUT and FBUT (ICC = 0.67 and 0.60 for averaged and first measurements, respectively). Bland-Altman analysis showed that the 95% limits of agreement for mNIBUT-aNIBUT and FBUT-aNIBUT were wider than those for FBUT-mNIBUT. The lack of agreement did not change when averaging the three measurements. Results from the OSDI indicated that most participants did not report symptoms. No significant correlations were observed between the OSDI total score and any of the tear metrics (p > 0.05). The aNIBUT demonstrated the best agreement with symptoms in this young and largely asymptomatic population. Tear break-up time measurements obtained by novice examiners showed limited reliability and agreement when comparing traditional methods (FBUT and mNIBUT) and the aNIBUT. Examiner-related variability may influence the assessment of tear-film stability, although causal inferences regarding examiner experience cannot be established from this study design. Repeating measurements did not improve the reliability and agreement between tests when performed by novice examiners.