Search Results (478)

Amanita exitialis is a lethal mushroom species found in southern China. Its amatoxins can cause acute liver injury with a high case-fatality rate. However, reports combining toxin detection in clinical specimens with autopsy pathology remain limited. We conducted a retrospective analysis of A. exitialis poisoning events treated at Chuxiong Yi Autonomous Prefecture People’s Hospital from 2019 to 2024. Toxins were measured in collected mushrooms, patient blood, and urine. Clinical data included demographics, complications, laboratory parameters, and autopsy findings. Associations between a time-weighted urinary amatoxin exposure metric and laboratory indices were assessed. Ten poisoning incidents involving 27 individuals were identified, including five deaths. We collected 10 mushroom samples, 120 urine samples, and 108 blood samples. α-amanitin, β-amanitin, phallacidin, and phallisacin were detected in mushrooms and urine. The detection rates of α-AMA, β-AMA, PCD, and PSC in urine samples were 31.67%, 5.00%, 38.33%, and 49.17%, respectively. Only three blood samples tested positive for α-AMA. The time-weighted urinary amatoxin exposure metric was positively correlated with total bilirubin (TBIL), aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), creatinine (Cr), creatine kinase (CK), creatine kinase isoenzymes (CK-MB), prothrombin time (PT), activated partial thromboplastin time (APTT), and international normalized ratio (INR). Early symptoms included nausea, vomiting, diarrhea, abdominal pain, and distention; later findings involved injury to the liver, kidneys, intestines, heart, and lungs. On the fourth day following ingestion, there was a marked increase in bilirubin levels and a concurrent decrease in liver enzymes, indicating severe damage to the hepatocytes. Platelet count, white blood cell count, hemoglobin, and red blood cell count decreased over time. Autopsies demonstrated hepatic, renal, and myocardial injury, gastrointestinal mucosal exfoliation, and multiorgan hemorrhage. In summary, A. exitialis poisoning is primarily characterized by liver damage, accompanied by injuries to the kidneys, myocardium, and intestines, as well as multiorgan hemorrhaging, which may lead to blood toxicity. The detection rate of toxins in urine samples is relatively high, and early urine toxin testing can help clarify the diagnosis and guide treatment. Full article

Background: The citrus processing industry generates over 40 million tons of waste annually, representing a significant environmental challenge. Citrus by-products are rich in bioactive compounds with proven health benefits. This study aims to upcycle citrus waste by developing green extracts and evaluating their biological activities for cosmeceutical applications. Methods: Three NaDES formulations—choline chloride–urea (ChCl: U), choline chloride–citric acid (ChCl: CA), and betaine–urea (Bet: U)—were optimized to extract polyphenols from orange and lemon waste using roller agitation. Extracts were characterized by HPLC–ESI–MS/MS. Biological activities were assessed in human keratinocytes (HaCaT). Antioxidant activity was measured using a chemiluminescent assay that detects intracellular H₂O₂ production. The wound-healing potential was evaluated using scratch assays, and cytokine release (IL-6, IL-8, IL-1β, IL-10) was assessed by ELISA. DNA damage protection was evaluated by quantifying 53BP1 foci following genotoxic exposure (neocarzinostatin). Results: All NaDES extracts showed high polyphenol content, with hesperidin being the primary compound. Pretreatment with the extracts for 24 h significantly reduced intracellular H₂O₂ levels, confirming their antioxidant efficacy. In scratch assays, extracts enhanced wound closure; notably, the Bet: U-derived orange extract achieved complete closure within 48 h. All extracts increased IL-6 and IL-8 release, consistent with an early pro-regenerative response. Pretreatment with the Bet: U orange extract lowered the number of cells with high 53BP1 foci after genotoxic stress, indicating partial DNA damage protection. Conclusions: These findings highlight citrus by-product extracts as sustainable bioactive ingredients with great potential for skin repair and anti-aging formulations, promoting responsible cosmeceutical innovation. Full article

In₂O₃ nanoparticles were obtained by annealing precursors that had been hydrothermally/solvothermally synthesized at 200 °C using In(NO₃)₃·4.5H₂O as the starting material. Three solvents were used for the synthesis, namely water, alcohol and ethylene glycol. Urea or glycine additives were introduced into the reaction mixtures as stabilizing and structure-forming agents. The nanopowders obtained were characterized using X-ray diffraction, scanning and transmission electron microscopy, low-temperature nitrogen adsorption and X-ray photoelectron spectroscopy. The gas-sensing characteristics of the indium oxide-based sensors were investigated for the detection of hydrogen in air. It has been established that the nature of the solvent determines the phase composition and structure of indium oxide, while organic additives reduce the particle size and increase the specific surface area. It should be noted that the addition of glycine to an alcohol solution of indium nitrate during synthesis produces a phase transformation. The results show that the sensor based on In₂O₃ synthesized using this mixture has the best hydrogen sensing properties of all the materials considered in this study. Full article

Bicarbonate deficiency is a common complication of acute kidney injury (AKI) and chronic kidney disease (CKD), resulting from impaired renal bicarbonate handling. It promotes bone demineralization and calcium–phosphate (CaxP) imbalance. In dogs, elevated CaxP has been linked to increased frequency and severity of bicarbonate deficiency. This 10-year study evaluated the prevalence and degree of bicarbonate deficiency in 618 cats diagnosed with AKI, acute-on-chronic kidney disease (ACKD), and CKD, and its association with azotemia and mineral imbalance. A retrospective analysis was conducted on serum biochemical profiles, including creatinine, urea, ionized and total calcium, phosphate, CaxP, and bicarbonate. Cats with incomplete datasets or receiving sodium bicarbonate therapy were excluded. According to IRIS guidelines, bicarbonate deficiency was defined as <16 mmol/L and categorized as moderate (12–16 mmol/L) or severe (<12 mmol/L). Deficiency was found in 45% of cats, of which 63% were moderate and 37% severe. Prevalence was higher in AKI (58%) and ACKD (60%) compared to CKD (38%) (p = 0.002). Negative correlations between bicarbonate and creatinine, phosphate, and CaxP were detected in AKI and CKD, while in ACKD, only creatinine and phosphate correlated. Deficiency increased with disease severity in AKI (p = 0.0145) and CKD (p < 0.0001). Cats with CaxP > 70 mg²/dL² had higher deficiency rates (p < 0.0001). However, 21% with elevated CaxP had normal bicarbonate, suggesting mineral imbalance may occur independently of acidosis. Full article

Background/Objectives: Eleutherococcus senticosus (Siberian ginseng) is an adaptogenic plant widely recognized for its antioxidant and immunomodulatory properties; however its hepatoprotective potential properties are unexplored. This study aimed to evaluate whether the fruit extract of E. senticosus contains chemical constituents with hepatoprotective effects in a paracetamol-induced liver injury model in mice. Methods: Female BALB/c mice were randomized into five groups: control, paracetamol (300 mg/kg, IP), E. senticosus extract (750 or 1500 mg/kg, PO) + paracetamol, and silymarin (50 mg/kg) + paracetamol. Extracts were administered for seven days before paracetamol challenge. Biochemical markers (ALT, AST, urea, creatinine, protein, albumin) and hematological parameters were assessed, and organs were subjected to histopathological examination. Phytochemical characterization of the extract was performed using UHPLC-DAD-MS and ICP-OES. Results: The 750 mg/kg dose of E. senticosus extract maintained ALT, AST, urea, and creatinine levels close to control values, while the higher dose (1500 mg/kg) was less effective and showed an increase in serum urea. Both extract doses and silymarin attenuated creatinine elevation induced by paracetamol. No histopathological changes were detected in the kidneys or brains of treated animals. Phytochemical analysis revealed high contents of phenolic acids (chlorogenic and dicaffeoylquinic acids), flavonoids, amino acids, and essential minerals. Conclusions: E. senticosus fruit extract demonstrated a hepatoprotective effect at an optimal dose (750 mg/kg), indicating a potential dose-dependent effect. The absence of histopathological alterations in key organs supports the fruit extract’s safety. Full article

A simplified kinetic model for the quantitative analysis of a potentiometric, pH-based urea biosensor is presented. The device was an electrolyte–insulator–semiconductor capacitor (EISCAP) with a pH-sensitive Ta₂O₅ gate functionalized by a polyallylamine hydrochloride (PAH)/urease bilayer. Within the steady-state approximation, the kinetic equations yielded an implicit algebraic relation linking the bulk urea concentration to the local pH at the sensor surface. Numerical solution of this equation, combined with a fitting routine, provides the apparent Michaelis–Menten constant ($K_M$) and the normalized maximum reaction rate (${\overline{k}}_V$). Validation against the literature data confirmed the reliability of the approach. Experimental results were then analyzed in both phosphate buffer (PBS) and artificial urine (AU), covering urea concentrations of 0.1–50 mM. The fitted parameters showed comparable $K_M$ values of 10.9 mM (PBS) and 32.4 mM (AU), but strongly different ${\overline{k}}_V$ values: $2.2\times {10}^{-4}$ (PBS) versus $8.6\times {10}^{-7}$ (AU). The three-order reduction in AU was attributed to the inhibitory effects inherent to complex biological fluids. These findings highlight the importance of the model-based quantitative analysis of EISCAP biosensors, enabling the accurate characterization of immobilized enzyme layers and guiding optimization for applications in realistic sample matrices. Full article

Falciparum malaria is a life-threatening vector-borne disease prevalent in tropical and subtropical regions. The complexity of severe malaria demands a thorough investigation of host–parasite interactions. Twenty male Sprague Dawley rats were divided into two groups: uninfected controls and Plasmodium berghei-infected rats, infected via intraperitoneal injection of parasitized red blood cells. Serum samples were analysed using high-resolution untargeted Gas Chromatography–Time-of-Flight Mass Spectrometry. Metabolomic analyses revealed altered metabolites and enriched metabolic pathways. Distinct metabolite profiles were observed between infected and control groups. Infected rats showed elevated urea levels and reduced concentrations of 1,5-anhydroglucitol, D-(+)-Talose, and arachidonic acid. Pathway analysis revealed significant enrichment of the glucose-alanine cycle, alpha-linolenic acid metabolism, and linoleic acid metabolism in infected rats. Minimal enrichment was observed in arachidonic acid metabolism and lactose biosynthesis. The upregulation of the glucose-alanine cycle suggests increased gluconeogenesis in response to parasite-induced glucose depletion and energy demand. Elevated urea indicates enhanced amino acid catabolism. These findings highlight the potential of metabolomics as a diagnostic tool for malaria detection and prognosis. Full article

This study presents a novel ratiometric fluorescent sensor based on Förster resonance energy transfer (FRET) between glutathione (GSH)-coated CdTe quantum dots (CdTe/GSH QDs) and bovine serum albumin (BSA)-coated Au nanoclusters (AuNCs/BSA) for dopamine (DA) detection. The nanoparticles were characterized using transmission electron microscopy (TEM), zeta potential measurements, Fourier transform infrared (FTIR) spectroscopy, UV-Vis absorption and fluorescence spectroscopy. Key FRET parameters, including energy transfer efficiency (E), donor–acceptor distance (r), Förster distance (R₀), and the overlap integral (J), were determined. The interactions between the CdTe/GSH-AuNCs/BSA conjugate and DA were investigated, revealing a dual mechanism of QDs fluorescence quenching that involves both energy and electron transfer. The average lifetime values and spectral profiles of CdTe/GSH QDs, both in the absence and presence of DA, suggest a dynamic fluorescence quenching process. The variation in the ratiometric signal with increasing DA concentration demonstrated a linear response within the range of 0–250 µM, with a correlation coefficient of 0.9963 and a detection limit of 6.9 nM. This proposed nanosensor exhibited selectivity against potential interfering substances, including urea, glucose, BSA, GSH, citric acid, and metal ions such as Na⁺ and Ca²⁺. The conjugate also demonstrates excellent cytocompatibility and enhances cell proliferation in HeLa epithelial cells, making it suitable for biological applications. It was successfully employed for DA detection in urine samples, achieving recoveries ranging from 99.1% to 104.2%. The sensor is highly sensitive, selective, rapid, and cost-effective, representing a promising alternative for DA detection across various sample types. Full article

Postmortem diagnosis of sudden cardiac death (SCD) may escape detection due to the absence of thrombi and slow development of structural and immunohistochemical changes. Therefore, this study explores the possibility of analyzing relevant clinical chemistry biomarkers in myocardial homogenates and serum. Following an initial pilot study, myocardial samples from 113 autopsy cases were homogenized with distilled water, T-PER or 2 M urea. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK-MB), lactate dehydrogenase (LDH), orosomucoid and total protein were analyzed with an IndikoPlus and a subset was also analyzed with a Roche Cobas 8000 c701 analyzer, which also provided results for cardiac Troponin T, myoglobin and NT-proBNP. Although the yields varied with different extraction buffers depending on the analyte, distilled water was often as effective as T-PER and 2 M urea extraction for most analytes. Biomarker levels were consistently higher in the myocardial homogenates than in serum. Proteomic profiling on a subset confirmed higher concentrations of the cardiac markers in the tissue samples than in serum. Finally, we investigated whether selected markers could support the diagnosis of acute cardiac disease by classifying cases as sudden cardiac death (SCD) or controls. There was no significant difference in serum concentrations of the selected biomarkers between SCD cases and controls, whereas a significant loss of several markers was observed in SCD myocardial samples as compared to controls. Hence, our results suggest that analysis of tissue homogenates is likely better for detecting early ischemia, and we show that an in-house benchtop multi-analyzer can provide rapid results to assist the pathologist’s decision-making during autopsy. Full article

Background/Objectives: Urinary angiotensin-converting enzyme (uACE) activity has long been regarded as a promising biomarker for kidney and cardiovascular diseases; however, its clinical applicability has been limited by the presence of endogenous urinary inhibitors and technically demanding assay protocols. We aimed to establish a fast and reproducible method for measuring uACE activity to identify the inhibitory compounds responsible for previous assay failures and to define practical preanalytical conditions suitable for routine laboratory implementation. Methods: A fluorescence-based kinetic assay was optimized for urine samples. Endogenous inhibitors were isolated by membrane filtration and chemically characterized, while the effect of sample dilution was evaluated as a simplified alternative for eliminating inhibitory interference. We assessed the stability of ACE activity under various storage conditions to support reliable measurement. Results: Urea (IC₅₀ = 1.18 M), uric acid (IC₅₀ = 3.61 × 10⁻³ M), and urobilinogen (IC₅₀ = 2.98 × 10⁻⁴ M) were identified as the principal reversible inhibitors, jointly accounting for up to 90% suppression of uACE activity. Their inhibitory effect was effectively eliminated by a 128-fold dilution. ACE activity remained stable for 24 h at 25 °C but was completely lost after freezing. A strong positive correlation between uACE activity and creatinine concentration (r = 0.76, p < 0.0001) justified normalization. ACE activity-to-creatinine ratio turned out to be significantly lower in ACE inhibitor-treated patients than in untreated controls (6.49 vs. 36.69 U/mol, p < 0.0001). Conclusions: Our findings demonstrate that accurate measurement of uACE activity is feasible using a rapid dilution-based protocol. The normalized ACE activity can serve as a practical biomarker for detecting pharmacological ACE inhibition and monitoring therapy adherence in cardiovascular care and may also provide insight into renal pathophysiology such as tubular injury or local RAAS-related processes. Full article

Hyperuricemia, characterized by elevated serum uric acid levels, is a major risk factor for gout and kidney disease. This study evaluated the antihyperuricemic effects of Cornus officinalis extract (COE) using urate transporter 1 (URAT1)-expressing oocytes and a hyperuricemia rat model. COE effectively inhibited uric acid absorption by modulating URAT1, with an IC₅₀ value of 3.24 µg/mL. In the hyperuricemia model, COE administration (100 and 200 mg/kg) significantly reduced serum uric acid levels and increased urinary uric acid excretion. The primary constituents of COE, morroniside (MO) and loganin (LO) exerted similar effects, with MO exhibiting potent inhibition of uric acid absorption even at low concentrations. Kidney tissue analysis revealed a reduction in blood urea nitrogen (BUN) levels, indicating improved renal function. Liver function parameters (ALT, AST, and LDH) remained unchanged, suggesting an absence of hepatotoxicity. Ultra-high-performance liquid chromatography with charged aerosol detection (UHPLC-CAD) analysis identified MO (17.8 mg/g), LO (9.8 mg/g), and cornin (1.4 mg/g) as the principal components of COE. These findings suggest that COE enhances uric acid excretion via URAT1 regulation and exerts renoprotective effects, highlighting its potential as an antihyperuricemic agent. Furthermore, MO and LO were identified as the primary active constituents, and COE appears to be a promising therapeutic candidate with a favorable safety profile. Full article

In 40 unique pediatric biopsy samples, this study aimed to compare the obtained results of Helicobacter pylori DNA detection using next-generation sequencing (NGS), a real-time PCR-based IVD-certified kit and an established high resolution melting real-time PCR-based method for H. pylori-specific ureA gene. From the same group, the H. pylori DNA was identified in 16 (40.0%) samples in both real-time PCR-based methods, with quantification cycle (Cq) values ranging from 17.51 to 32.21 for the IVD kit. NGS was able to detect H. pylori DNA in 14 (35.0%) samples, with read counts between 7768 and 42,924. While all three methods showed similar detection rates, both PCR variants were slightly more sensitive, identifying H. pylori in two additional samples not detected by NGS. The study highlights the strengths and limitations of each method. NGS, though promising due to its high sensitivity and ability to detect low bacterial load, is still limited by its cost and complexity. Despite these challenges, NGS could complement PCR in diagnosing difficult or ambiguous cases, enabling the detection of multiple pathogens simultaneously. Especially when other infectious etiologies are suspected, NGS could be considered, though PCR variants remain a more attractive and cost-effective option for routine H. pylori detection. Full article

A nanoliter-scale fabrication method was applied to construct a colorimetric lateral flow strip for urea detection (Urea-CLFS). The device involves two main papers: a nitrocellulose membrane (NC-Mb) for urease enzyme immobilization and chromatography paper (CH-PP) containing a phenol red indicator. Urea-CLFS is a tool for detecting urea that is based on enzyme catalysis and the change in color of phenol red when urea is present. The Urea-CLFS fabrication was made possible by the minimal amount of nanoliters used in reagent consumption. The use of small arrays of phenol red dots provides a higher response result compared to single dots applied on CH-PP. To find the most effective design, it analyzed how urease was aligned on NC-Mb horizontally and vertically. According to our findings, the vertical alignment of the urease enzyme on NC-Mb leads to a prolonged reaction time, which leads to higher product production. The optimization process included optimizing various parameters, including the layer number of phenol red on CH-PP, phenol red concentration, urease concentration, reaction time, and sample volume. Under optimal conditions, the Urea-CLFS provided a linear range of 0.25–8.0 mmol L⁻¹ with an LOD of 0.34 mmol L⁻¹, which is sufficient for human health diagnostics. The accuracy of the Urea-CLFS was demonstrated by the recovery of the human urine sample between 95 ± 3% and 103 ± 3% (n = 3). Full article

Typical fertilizer applicators are often restricted in performance due to non-uniformity in distribution, required labor and time intensiveness, high discharge rate, chemical input wastage, and fostering weed proliferation. To address this gap in production agriculture, an automated variable-rate fertilizer applicator was developed for the cotton crop that is based on deep learning-initiated electronic control unit (ECU). The applicator comprises (a) plant recognition unit (PRU) to capture and predict presence (or absence) of cotton plants using the YOLOv7 recognition model deployed on-board Raspberry Pi microprocessor (Wale, UK), and relay decision to a microcontroller; (b) an ECU to control stepper motor of fertilizer metering unit as per received cotton-detection signal from the PRU; and (c) fertilizer metering unit that delivers precisely metered granular fertilizer to the targeted cotton plant when corresponding stepper motor is triggered by the microcontroller. The trials were conducted in the laboratory on a custom testbed using artificial cotton plants, with the camera positioned 0.21 m ahead of the discharge tube and 16 cm above the plants. The system was evaluated at forward speeds ranging from 0.2 to 1.0 km/h under lighting levels of 3000, 5000, and 7000 lux to simulate varying illumination conditions in the field. Precision, recall, F1-score, and mAP of the plant recognition model were determined as 1.00 at 0.669 confidence, 0.97 at 0.000 confidence, 0.87 at 0.151 confidence, and 0.906 at 0.5 confidence, respectively. The mean absolute percent error (MAPE) of 6.15% and 9.1%, and mean absolute deviation (MAD) of 0.81 g/plant and 1.20 g/plant, on application of urea and Diammonium Phosphate (DAP), were observed, respectively. The statistical analysis showed no significant effect of the forward speed of the conveying system on fertilizer application rate (p > 0.05), thereby offering a uniform application throughout, independent of the forward speed. The developed fertilizer applicator enhances precision in site-specific applications, minimizes fertilizer wastage, and reduces labor requirements. Eventually, this fertilizer applicator placed the fertilizer near targeted plants as per the recommended dosage. Full article

Hypervirulent Klebsiella pneumoniae (hvKp) has emerged as a significant public health concern, yet rare sublineages remain poorly characterized. Here, we described a K30-ST198 hvKp sublineage identified in four isolates from two patients, including three sequential strains (K30B1, K30B2, K30B3) recovered over eight months from recurrent liver abscesses and one strain (K30-HUMV1) from a urinary tract infection. All isolates exhibited a yYpermucoviscous phenotype and resistance restricted to ampicillin and amoxicillin. Screening with the eazyplex hvKp assay detected ybt and rmpA in all strains, yielding a virulence score of 1. Biofilm production was strong in K30B1, K30B2, moderate in K30-HUMV1, but weak in K30B3. In the Galleria mellonella infection model, K30B1 showed higher virulence than the other isolates. Whole-genome sequencing identified the ICEKp1 carrying hypervirulence-associated genes (ybt, pagO, rmpAC, iroBCDN) together with additional virulence factors (fim, mrkD, uge, ureA, wabG, wcaJ, mliC), while antibiotic resistance genes were limited to fosA and bla_SHV-77. Protein structures and their functional domains were predicted using AlphaFold v3.0.1 and ColabFold v1.5.5, based on pLDDT scores, providing further insights into gene functionality. This work represents one of the first detailed characterizations of K30-ST198 hvKp, underscoring the need for integrated genomic, phenotypic, and structural approaches in hvKp surveillance. Full article