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14 pages, 1635 KB  
Article
Toxic Effects of Waterborne Nitrite on LC50, Hematological Parameters, and Plasma Biochemistry in Starry Flounder (Platichthys stellatus)
by Bijae Gong, Hyeong Su Kim, Cheol Young Choi, Sung-Pyo Hur and Jun-Hwan Kim
Toxics 2025, 13(9), 748; https://doi.org/10.3390/toxics13090748 - 2 Sep 2025
Abstract
Nitrite is a common environmental pollutant in aquaculture systems, where high levels can severely impair fish physiology and survival. This study aimed to evaluate the acute toxicity of waterborne nitrite in starry flounder (Platichthys stellatus). Fish (mean weight 145.69 ± 16.06 [...] Read more.
Nitrite is a common environmental pollutant in aquaculture systems, where high levels can severely impair fish physiology and survival. This study aimed to evaluate the acute toxicity of waterborne nitrite in starry flounder (Platichthys stellatus). Fish (mean weight 145.69 ± 16.06 g, mean total length 22.78 ± 0.70 cm) were exposed to nitrite concentrations of 0, 25, 50, 100, 200, 400, and 800 mg NO2/L for 96 h. The lethal concentration 50 (LC50) of nitrite for P. stellatus was determined to be 574.47 mg NO2/L. Hematological parameters such as red blood cell counts (RBCs), hemoglobin (Hb), and hematocrit (Hct) were significantly decreased by nitrite exposure. Plasma components including calcium (Ca2+), glucose, cholesterol, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were significantly changed by nitrite exposure. The results of this study suggest that acute exposure to waterborne nitrite (>200 mg NO2/L) adversely affects survival rates, hematological parameters, and plasma components in P. stellatus. These findings provide important baseline data for nitrite toxicity assessment in P. stellatus. Full article
(This article belongs to the Section Exposome Analysis and Risk Assessment)
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14 pages, 3607 KB  
Article
Association of Erythrocyte-Related Indices with Immune-Related Adverse Events and Survival of Lung Cancer Patients Receiving Immune Checkpoint Inhibitors
by Zhan Wang, Ting Zou, Chen-Wei Liao, Xiang-Ping Li, Zhao-Qian Liu, Ze-Fu Liu and Juan Chen
Pharmaceuticals 2025, 18(9), 1299; https://doi.org/10.3390/ph18091299 - 29 Aug 2025
Viewed by 99
Abstract
Background: Lung cancer has the highest lethality rate among malignancies worldwide. Immunotherapy is one of the common treatments for lung cancer patients. There are two main types of immunotherapies: one targets programmed cell death 1 (PD-1), and the other targets programmed cell [...] Read more.
Background: Lung cancer has the highest lethality rate among malignancies worldwide. Immunotherapy is one of the common treatments for lung cancer patients. There are two main types of immunotherapies: one targets programmed cell death 1 (PD-1), and the other targets programmed cell death ligand 1 (PD-L1). These two belong to the class of immune checkpoint inhibitors (ICIs). However, immune-related adverse reactions (irAEs) were the main reasons affecting its clinical therapeutic effect. Methods: This retrospective cohort study analyzed red blood cell count (RBC), hematocrit (HCT), erythrocyte mean corpuscular volume (MCV) and immunotherapy outcomes in 920 lung cancer patients receiving immunotherapy from April 2019 to May 2023. Results: We found that high levels of RBC (>4.105 × 109, p = 0.007, OR = 0.467, 95%CI: 0.268~0.812) and MCV (>86.35, p = 0.017, OR = 0.0.441, 95%CI: 0.224~0.865) were significantly related to the better response of ICIs immunotherapy in patients. Patients with high levels of HCT (>39.75%, p = 0.035, OR = 0.737, 95%CI: 0.555~0.979) may have a lower rate of irAEs occurrence. Meanwhile, patients with a low level of RBCs (≤4.635 × 109, p < 0.001, OR = 1.636, 95%CI: 1.365~1.960) may have a longer period of PFS (progression-free survival), and patients with RBC (≤4.43 × 109, p = 0.033, OR = 0.480, 95%CI: 0.244~0.941) may have a longer time of OS (overall survival). Conclusions: The findings indicate that the levels of RBC, MCV and HCT were significantly associated with the response and irAEs of ICIs in lung cancer patients. The levels of RBC might act as a possible biomarker for predicting the survival of lung cancer patients who are receiving ICI therapy. Full article
(This article belongs to the Special Issue Adjuvant Therapies for Cancer Treatment: 2nd Edition)
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21 pages, 3101 KB  
Article
Filling a Gap in Quercus Phylogeny: Molecular Phylogenetic Evidence, Morphometric and Biogeographic History of Quercus petraea subsp. pinnatiloba Matt. Liebl from Türkiye
by Pelin Acar
Diversity 2025, 17(9), 599; https://doi.org/10.3390/d17090599 - 26 Aug 2025
Viewed by 244
Abstract
Quercus petraea subsp. pinnatiloba is a narrowly distributed oak taxon in southeastern Türkiye, and its taxonomic position has long remained uncertain. This study aims to clarify its distinctiveness by integrating morphological, molecular, and biogeographical evidence. Principal Component Analysis (PCA) and Stepwise Discriminant Analysis [...] Read more.
Quercus petraea subsp. pinnatiloba is a narrowly distributed oak taxon in southeastern Türkiye, and its taxonomic position has long remained uncertain. This study aims to clarify its distinctiveness by integrating morphological, molecular, and biogeographical evidence. Principal Component Analysis (PCA) and Stepwise Discriminant Analysis (SDA) of 14 leaf traits revealed that subsp. pinnatiloba constitutes a morphologically stable and distinctly differentiated group from other Q. petraea subspecies and closely related taxa, characterized by key diagnostic traits such as petiole length (PL), lamina length (LL), length of leaf blade at its broadest point (WP), and lobe width at the tip of the widest lobe (LW). Phylogenetic analyses based on nuclear ITS and plastid markers (rbcL, psbA-trnH) confirmed its placement within sect. Quercus, yet consistently distinguished it genetically from other subspecies for the first time. Molecular dating (BEAST) suggested divergence in the Miocene (11 Mya with 95% HPD 3.01, 20.95) while RASP biogeographical analysis indicated an origin in the Euro-Siberian region with later dispersal into the Mediterranean. These integrative results support its recognition at species rank as Quercus pinnatiloba, clarifying its phylogenetic placement and underscoring the conservation importance of this lineage. Full article
(This article belongs to the Section Phylogeny and Evolution)
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18 pages, 3836 KB  
Article
A Microfluidic Approach for Assessing the Rheological Properties of Healthy Versus Thalassemic Red Blood Cells
by Hao Jiang, Xueying Li, Zhuoyan Liu, Siyu Luo, Junbin Huang, Chun Chen, Rui Chen and Fenfang Li
Micromachines 2025, 16(8), 957; https://doi.org/10.3390/mi16080957 - 19 Aug 2025
Viewed by 445
Abstract
The deformability of red blood cells (RBCs) is critical for microvascular circulation and is impaired in hematological disorders such as thalassemia, a prevalent public health concern in Guangdong, China. While microfluidics enable high-precision deformability assessment, current studies lack standardization in deformation metrics and [...] Read more.
The deformability of red blood cells (RBCs) is critical for microvascular circulation and is impaired in hematological disorders such as thalassemia, a prevalent public health concern in Guangdong, China. While microfluidics enable high-precision deformability assessment, current studies lack standardization in deformation metrics and rarely investigate post-deformation recovery dynamics. This study introduces an automated microfluidic platform for systematically evaluating RBC deformability in healthy and thalassemic individuals. A biomimetic chip featuring 4 µm, 8 µm, and 16 µm wide channels (7 µm in height) was designed to simulate capillary dimensions, with COMSOL CFD numerical modeling validating shear stress profiles. RBC suspensions (107 cells/mL in DPBS) were hydrodynamically focused through constrictions while high-speed imaging (15,000 fps) captured deformation–recovery dynamics. Custom-built algorithms with deep-learning networks automated cell tracking, contour analysis, and multi-parametric quantification. Validation confirmed significantly reduced deformability in Paraformaldehyde (PFA)-treated RBCs compared to normal controls. Narrower channels and higher flow velocities amplified shear-induced deformations, with more deformable cells exhibiting faster post-constriction shape recovery. Crucially, the platform distinguished thalassemia patient-derived RBCs from healthy samples, revealing significantly lower deformability in diseased cells, particularly in 4 µm channels. These results establish a standardized, high-throughput framework for RBC mechanical characterization, uncovering previously unreported recovery dynamics and clinically relevant differences in deformability in thalassemia. The method’s diagnostic sensitivity highlights its translational potential for screening hematological disorders. Full article
(This article belongs to the Special Issue Application of Microfluidic Technology in Bioengineering)
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24 pages, 5298 KB  
Article
Therapeutic Potential of Ozonated Ocimum basilicum L. from Saudi Arabia: Phytochemical Characterization and Enhanced Bioactivities
by Husam Qanash, Sulaiman A. Alsalamah, Abdulrahman S. Bazaid, Mohammed Ibrahim Alghonaim, Amro Duhduh and Ibtisam Hudani
Pharmaceuticals 2025, 18(8), 1223; https://doi.org/10.3390/ph18081223 - 19 Aug 2025
Viewed by 413
Abstract
Background/Objectives: Medicinal plants are an abundant source of bioactive molecules, particularly in arid environments, such as Saudi Arabia, where Ocimum basilicum L. (Saudi basil) has long been used for its therapeutic properties. This study aimed to examine the phytochemical profile and bioactivities [...] Read more.
Background/Objectives: Medicinal plants are an abundant source of bioactive molecules, particularly in arid environments, such as Saudi Arabia, where Ocimum basilicum L. (Saudi basil) has long been used for its therapeutic properties. This study aimed to examine the phytochemical profile and bioactivities of non-ozonated (untreated) and ozonated methanolic extracts of O. basilicum and to determine whether ozonation enhances their biological effects, with a focus on antidiabetic, anti-Alzheimer, anti-inflammatory, antimicrobial, and cytotoxic properties. Methods: Fresh leaves of O. basilicum were extracted with methanol, subjected to ozonation, and analyzed by HPLC. In vitro assays were conducted to evaluate α-amylase, α-glucosidase, and BChE inhibition, RBC membrane stabilization, antibacterial activity against Helicobacter pylori and cytotoxicity using normal lung fibroblasts (WI-38) and human colorectal adenocarcinoma cell line (Caco-2). Results: Ozonation modified the phytochemical profile, enriching chlorogenic and rosmarinic acids. Ozonated extracts exhibited stronger inhibition of α-amylase with an IC50 of 5.09 µg/mL compared to 13.6 µg/mL of untreated Saudi basil and α-glucosidase (IC50 6.15 µg/mL vs. 9.42 µg/mL). They also showed enhanced BChE inhibition with an IC50 of 13.4 µg/mL compared to 31.8 µg/mL of non-ozonated extract. In addition, ozonated extracts produced significant anti-inflammatory effects by stabilizing RBCs, with an IC50 of 8.04 µg/mL compared to 8.44 µg/mL for untreated extracts and 4.41 µg/mL for indomethacin. Ozonated extracts produced larger H. pylori inhibition zones (26.7 mm) and an MBC/MIC ratio of 1. Cytotoxicity testing revealed that ozonated extracts were less toxic to WI-38 cells, with IC50 values of 437.89 µg/mL versus 191.06 µg/mL, and 149.14 µg/mL compared to 103.7 µg/mL of untreated Saudi basil in Caco-2 cells. Conclusions: Ozonation enriches the phytochemical composition of O. basilicum, enhancing antidiabetic, neuroprotective, anti-inflammatory, and antibacterial activities while reducing cytotoxicity on normal cells. These findings support the potential of ozonated O. basilicum as a safe and promising natural therapeutic candidate for metabolic, neurodegenerative, and infectious diseases. Full article
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17 pages, 3914 KB  
Article
Green Synthesis of Chitosan-Coated Selenium Nanoparticles for Paclitaxel Delivery
by Mouhaned Y. Al-Darwesh, Maroua Manai, Hammouda Chebbi and Axel Klein
Nanomaterials 2025, 15(16), 1276; https://doi.org/10.3390/nano15161276 - 18 Aug 2025
Viewed by 485
Abstract
Selenium nanoparticles (Se NPs) were synthesized from Na2SeO3 using Foeniculum vulgare (fennel) seed extract as mild sustainable reductant, coated with chitosan (Ch), and loaded with Paclitaxel (PTX). The PTX release from the Se@Ch–PTX NPs and their cytotoxicity against MDA-MB-231 breast [...] Read more.
Selenium nanoparticles (Se NPs) were synthesized from Na2SeO3 using Foeniculum vulgare (fennel) seed extract as mild sustainable reductant, coated with chitosan (Ch), and loaded with Paclitaxel (PTX). The PTX release from the Se@Ch–PTX NPs and their cytotoxicity against MDA-MB-231 breast cancer cells was studied in view of an application as drug delivery platform. Thermogravimetric analysis (TGA) showed the thermal stability of the NPs up to 300 °C. UV–vis absorption and Fourier transform IR (FT-IR) spectroscopy allowed to trace surface species originating from the F. vulgare extract on the Se NPs, while the surface of the Se@Ch–PTX NPs is characterized from Ch and PTX functionalities. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed approximate spherical shaped NPs with sizes ranging from 10 to 40 nm. Zeta potential measurements showed a clear distinction between the −39 mV found the Se NPs and +57 mV for the Ch–PTX coated NPs. The NPs showed good biocompatibility with red blood cells (RBCs) in hemolytic activity assays, exhibiting no hemolytic effects at concentrations ranging from 50 to 400 µg/mL. In vitro release studies showed a sustained and pH-responsive release pattern with a maximum release of about 80% within 22 h for Se@Ch–PTX at pH = 3.5. The Se@Ch–PTX NPs showed high antiproliferative activity against MDA-MB-231 cells with an IC50 value of 12.3 µg/mL compared to about 36 for PTX and 52 µg/mL for the Se NPs. The reactive oxygen species (ROS) activity as studied through DPPH scavenging showed higher values for the Se@Ch–PTX NPs compared to the Se NP. Full article
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24 pages, 7673 KB  
Article
Development, Molecular Docking, and Anti-Anemia Potential of Polyherbal Formulation
by Deepak Bharati, Sakshi Nirhali, Abhijeet Puri, Popat Mohite and Sudarshan Singh
Biology 2025, 14(8), 1052; https://doi.org/10.3390/biology14081052 - 14 Aug 2025
Viewed by 389
Abstract
Anemia remains a widespread public health concern, and the search for interventions demonstrating potent anti-anemic activity is critical for reducing its impact among high-risk populations. Conventional iron therapies are associated with several complications and potential adverse effects. This study explored a polyherbal approach [...] Read more.
Anemia remains a widespread public health concern, and the search for interventions demonstrating potent anti-anemic activity is critical for reducing its impact among high-risk populations. Conventional iron therapies are associated with several complications and potential adverse effects. This study explored a polyherbal approach to develop a safer and more effective alternative treatment for anemia. A molecular docking study was initially performed to screen and evaluate alizarin, catechin, kaempferol, recesmol, rubiadin, and rutin, which are known for their antioxidant and hematinic potential. Using AutoDock Vina, these compounds were docked against the target protein (PDB ID: 6MOE) with EPE and ferrous ions as controls. Rutin demonstrated the highest binding affinity of −6.4 kcal/moL, whereas alizarin and rubiadin both followed closely with −6.3 kcal/moL, while kaempferol and ellagic acid exhibited a binding affinity of −6.2 kcal/moL. In comparison, the reference compounds tested ferrous ions, and native ligand EPE (−5.0 kcal/moL) and iron (−4.8 kcal/moL), showed mild affinities. Moreover, the tested compounds demonstrated stable binding, suggesting their potential relevance in modulating anemia-related pathways. Based on the docking results and traditional therapeutic values, a polyherbal formulation (PHF) was developed using methanolic extracts of Trigonella foenum-graecum, Emblica officinalis, Pterocarpus marsupium, Withania somnifera, Asparagus racemosus, Zingiber officinale, Rubia cordifolia, Boerhavia diffusa, and Adhatoda vasica. Phytochemical screening via HPTLC analysis was used to quantify the presence of gallic and ellagic acids. In addition, PHF showed significant antioxidant potential (DPPH IC50: 14.29 µg/mL; FRAP IC50: 58.57 µg/mL) and iron content (98.47 ppm) values. Furthermore, in vivo evaluation using a phenylhydrazine-induced hemolytic anemia model in Sprague Dawley rats revealed that the PHF achieved complete restoration of RBCs (6.15 ± 0.04), hemoglobin (14.82 ± 0.03 g/dL), and hematocrit (43.08 ± 0.28%) in anemic rats and improved histopathological features in the liver, spleen, and bone marrow. These results demonstrate that combined molecular and pharmacological evidence support the efficacy of PHF as a promising candidate for the management of anemia by enhancing erythropoiesis, improving iron metabolism, and reducing oxidative stress. Full article
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24 pages, 4213 KB  
Article
Influence of Morus alba Leaves Extract on Human Erythrocytes
by Stefano Putaggio, Annamaria Russo, Giuseppe Tancredi Patanè, Antonella Calderaro, Santa Cirmi, Ivana Verboso, Giuseppina Laganà, Silvana Ficarra, Davide Barreca, Françisco Raymo and Ester Tellone
Biology 2025, 14(8), 1005; https://doi.org/10.3390/biology14081005 - 5 Aug 2025
Viewed by 335
Abstract
Morus alba L. (MA) is a member of the Moraceae family, known as “white mulberry”. Due to the high levels of bioactive compounds, mulberry plants can be considered a good source of nutrients and antioxidant compounds. Our study aims to analyze the effect [...] Read more.
Morus alba L. (MA) is a member of the Moraceae family, known as “white mulberry”. Due to the high levels of bioactive compounds, mulberry plants can be considered a good source of nutrients and antioxidant compounds. Our study aims to analyze the effect of MA extract leaves on erythrocytes, focusing on its action on metabolism and membrane integrity. The choice of erythrocytes as a study model is based on their metabolic simplicity and their easy availability. Cell viability, following exposure of the cells to the extract, was evaluated by hemolysis, methemoglobin, caspase 3 activity and flow cytofluorimetric analysis; in addition, the effect of the pretreatment with the MA was detected after incubation of erythrocytes with different stressors. The impact on cell metabolism was evaluated by measuring anion flux kinetics, ATP levels and phosphatase activity. The results obtained show a peculiar (double) effect of the extract, which, on the one hand, probably by exploiting its component with antioxidant properties, protects the cell membrane by accumulating on the bilayer. On the other hand, the alteration of anion exchange could lead to the triggering of apoptosis and consequent cell death. The hypotheses, although excluded by our data, all point toward a beneficial and protective action of the extract on the health and vitality of RBCs. Full article
(This article belongs to the Section Biochemistry and Molecular Biology)
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29 pages, 10502 KB  
Article
A Comparative Bioinformatic Investigation of the Rubisco Small Subunit Gene Family in True Grasses Reveals Novel Targets for Enhanced Photosynthetic Efficiency
by Brittany Clare Thornbury, Tianhua He, Yong Jia and Chengdao Li
Int. J. Mol. Sci. 2025, 26(15), 7424; https://doi.org/10.3390/ijms26157424 - 1 Aug 2025
Viewed by 353
Abstract
Ribulose bisphosphate carboxylase (RuBisCO) is the primary regulator of carbon fixation in the plant kingdom. Although the large subunit (RBCL) is the site of catalysis, RuBisCO efficiency is also influenced by the sequence divergence of the small subunit (RBCS). This project compared the [...] Read more.
Ribulose bisphosphate carboxylase (RuBisCO) is the primary regulator of carbon fixation in the plant kingdom. Although the large subunit (RBCL) is the site of catalysis, RuBisCO efficiency is also influenced by the sequence divergence of the small subunit (RBCS). This project compared the RBCS gene family in C3 and C4 grasses to identify genetic targets for improved crop photosynthesis. Triticeae/Aveneae phylogeny groups exhibited a syntenic tandem duplication array averaging 326.1 Kbp on ancestral chromosomes 2 and 3, with additional copies on other chromosomes. Promoter analysis revealed a paired I-box element promoter arrangement in chromosome 5 RBCS of H. vulgare, S. cereale, and A. tauschii. The I-box pair was associated with significantly enhanced expression, suggesting functional adaptation of specific RBCS gene copies in Triticaeae. H. vulgare-derived pan-transcriptome data showed that RBCS expression was 50.32% and 28.44% higher in winter-type accessions compared to spring types for coleoptile (p < 0.05) and shoot, respectively (p < 0.01). Molecular dynamics simulations of a mutant H. vulgare Rubisco carrying a C4-like amino acid substitution (G59C) in RBCS significantly enhanced the stability of the Rubisco complex. Given the known structural efficiency of C4 Rubisco complexes, G59C could serve as an engineering target for enhanced RBCS in economically crucial crop species which, in comparison, possess less efficient Rubisco complexes. Full article
(This article belongs to the Special Issue Molecular Genetics, Genomics and Breeding in Field Crops)
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36 pages, 3621 KB  
Review
Harnessing Molecular Phylogeny and Chemometrics for Taxonomic Validation of Korean Aromatic Plants: Integrating Genomics with Practical Applications
by Adnan Amin and Seonjoo Park
Plants 2025, 14(15), 2364; https://doi.org/10.3390/plants14152364 - 1 Aug 2025
Viewed by 703
Abstract
Plant genetics and chemotaxonomic analysis are considered key parameters in understanding evolution, plant diversity and adaptation. Korean Peninsula has a unique biogeographical landscape that supports various aromatic plant species, each with considerable ecological, ethnobotanical, and pharmacological significance. This review aims to provide a [...] Read more.
Plant genetics and chemotaxonomic analysis are considered key parameters in understanding evolution, plant diversity and adaptation. Korean Peninsula has a unique biogeographical landscape that supports various aromatic plant species, each with considerable ecological, ethnobotanical, and pharmacological significance. This review aims to provide a comprehensive overview of the chemotaxonomic traits, biological activities, phylogenetic relationships and potential applications of Korean aromatic plants, highlighting their significance in more accurate identification. Chemotaxonomic investigations employing techniques such as gas chromatography mass spectrometry, high-performance liquid chromatography, and nuclear magnetic resonance spectroscopy have enabled the identification of essential oils and specialized metabolites that serve as valuable taxonomic and diagnostic markers. These chemical traits play essential roles in species delimitation and in clarifying interspecific variation. The biological activities of selected taxa are reviewed, with emphasis on antimicrobial, antioxidant, anti-inflammatory, and cytotoxic effects, supported by bioassay-guided fractionation and compound isolation. In parallel, recent advances in phylogenetic reconstruction employing DNA barcoding, internal transcribed spacer regions, and chloroplast genes such as rbcL and matK are examined for their role in clarifying taxonomic uncertainties and inferring evolutionary lineages. Overall, the search period was from year 2001 to 2025 and total of 268 records were included in the study. By integrating phytochemical profiling, pharmacological evidence, and molecular systematics, this review highlights the multifaceted significance of Korean endemic aromatic plants. The conclusion highlights the importance of multidisciplinary approaches including metabolomics and phylogenomics in advancing our understanding of species diversity, evolutionary adaptation, and potential applications. Future research directions are proposed to support conservation efforts. Full article
(This article belongs to the Special Issue Applications of Bioinformatics in Plant Science)
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14 pages, 4594 KB  
Article
Short-Term Exposure to Tebuconazole Triggers Haematological, Histological and Biochemical Disturbances in Rainbow Trout (Oncorhynchus mykiss)
by Akif Er
Toxics 2025, 13(8), 630; https://doi.org/10.3390/toxics13080630 - 27 Jul 2025
Viewed by 540
Abstract
Tebuconazole (TBZ), a triazole-class fungicide widely used in agriculture, is frequently detected in aquatic environments due to runoff and leaching, where it poses a threat to non-target aquatic organisms. This study investigates the acute toxicity of TBZ on juvenile rainbow trout (Oncorhynchus [...] Read more.
Tebuconazole (TBZ), a triazole-class fungicide widely used in agriculture, is frequently detected in aquatic environments due to runoff and leaching, where it poses a threat to non-target aquatic organisms. This study investigates the acute toxicity of TBZ on juvenile rainbow trout (Oncorhynchus mykiss), a commercially important cold-water fish species. The 96 h LC50 value was determined to be 9.05 mg/L using probit analysis. In addition to mortality, the physiological responses of fish exposed to both LC50 and maximum tolerance concentration (MTC; 6 mg/L) were evaluated through haematological and histological assessments. TBZ exposure significantly suppressed key haematological parameters, particularly WBC, RBC, HGB, HCT, and LYM, indicating immunosuppression and potential hypoxia. Histological examination revealed progressive and regressive damage in gill tissues, including epithelial lifting, hyperplasia, and hypertrophy, which were more severe in the LC50 group. These alterations were quantified using a semi-quantitative scoring system. Additionally, significant changes in biochemical parameters such as ALT, AST, creatinine, total protein, and glucose levels were observed, further indicating hepatic and renal dysfunctions induced by TBZ exposure. The findings demonstrate that TBZ exposure induces substantial physiological and structural impairments in rainbow trout, highlighting the importance of assessing the ecological risks of fungicide contamination in aquatic environments. The study also provides a dose–response model that can be used to estimate mortality risk in aquaculture operations exposed to TBZ. Full article
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16 pages, 3978 KB  
Article
Cepharanthine Promotes Ca2+-Independent Premature Red Blood Cell Death Through Metabolic Insufficiency and p38 MAPK/CK1α/COX/MLKL/PKC/iNOS Signaling
by Shaymah H. Alruwaili, Jawaher Alsughayyir and Mohammad A. Alfhili
Int. J. Mol. Sci. 2025, 26(15), 7250; https://doi.org/10.3390/ijms26157250 - 27 Jul 2025
Viewed by 440
Abstract
Nonspecific toxicity to normal and malignant cells restricts the clinical utility of many anticancer drugs. In particular, anemia in cancer patients develops due to drug-induced toxicity to red blood cells (RBCs). The anticancer alkaloid, cepharanthine (CEP), elicits distinct forms of cell death including [...] Read more.
Nonspecific toxicity to normal and malignant cells restricts the clinical utility of many anticancer drugs. In particular, anemia in cancer patients develops due to drug-induced toxicity to red blood cells (RBCs). The anticancer alkaloid, cepharanthine (CEP), elicits distinct forms of cell death including apoptosis and autophagy, but its cytotoxicity to RBCs has not been investigated. Colorimetric and fluorometric techniques were used to assess eryptosis and hemolysis in control and CEP-treated RBCs. Cells were labeled with Fluo4/AM and annexin-V-FITC to measure Ca2+ and phosphatidylserine (PS) exposure, respectively. Forward scatter (FSC) was detected to estimate cell size, and extracellular hemoglobin along with lactate dehydrogenase and aspartate transaminase activities were assayed to quantify hemolysis. Physiological manipulation of the extracellular milieu and various signaling inhibitors were tested to dissect the underlying mechanisms of CEP-induced RBC death. CEP increased PS exposure and hemolysis indices and decreased FSC in a concentration-dependent manner with prominent membrane blebbing. Although no Ca2+ elevation was detected, chelation of intracellular Ca2+ by BAPTA-AM reduced hemolysis. Whereas SB203580, D4476, acetylsalicylic acid, necrosulfonamide, and melatonin inhibited both PS exposure and hemolysis, staurosporin, L-NAME, ascorbate, caffeine, adenine, and guanosine only prevented hemolysis. Interestingly, sucrose had a unique dual effect by exacerbating PS exposure and reversing hemolysis. Of note, blocking KCl efflux augmented PS exposure while aggravating hemolysis only under Ca2+-depleted conditions. CEP activates Ca2+-independent pathways to promote eryptosis and hemolysis. The complex cytotoxic profile of CEP can be mitigated by targeting the identified modulatory pathways to potentiate its anticancer efficacy. Full article
(This article belongs to the Special Issue Blood Cells in Human Health and Disease)
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11 pages, 3019 KB  
Article
DNA Metabarcoding Reveals Seasonal Variations in Crop-Foraging Behavior of Wild Rhesus Macaques (Macaca mulatta)
by Yun Wang, Hongjia Li, Gongyuan Shi, Heqin Cao, Manfang He and Haijun Su
Diversity 2025, 17(8), 517; https://doi.org/10.3390/d17080517 - 26 Jul 2025
Viewed by 326
Abstract
The ecological drivers of wildlife crop-foraging behavior—whether as a compensatory response to natural resource scarcity or as opportunistic exploitation of anthropogenic food sources—remain poorly understood in human–wildlife conflict research. Traditional methodologies, which primarily rely on direct observation and morphological identification, have limitations in [...] Read more.
The ecological drivers of wildlife crop-foraging behavior—whether as a compensatory response to natural resource scarcity or as opportunistic exploitation of anthropogenic food sources—remain poorly understood in human–wildlife conflict research. Traditional methodologies, which primarily rely on direct observation and morphological identification, have limitations in comprehensively quantifying wildlife dietary composition, particularly in accurately distinguishing between morphologically similar plant species and conducting precise quantitative analyses. This study utilized DNA metabarcoding technology (rbcL gene markers) to identify and quantify plant dietary components through fecal sample analysis, systematically investigating the dietary composition and patterns of agricultural resource utilization of wild rhesus macaques (Macaca mulatta) in human–wildlife interface zones of southwestern China. A total of 29 rhesus macaque fecal samples were analyzed (15 from spring and 14 from winter), identifying 142 plant genera, comprising 124 wild plant genera, and 18 crop genera. The results revealed distinct seasonal foraging patterns: crops accounted for 32.11% of the diet in winter compared to 7.66% in spring. Notably, rhesus macaques continued to consume crops even during spring when wild resources were relatively abundant, challenging the traditional hypothesis driven by resource scarcity and suggesting that crop-foraging behavior may reflect an opportunistic, facultative resource selection strategy. This study demonstrates the significant value of DNA metabarcoding technology in wildlife foraging behavior research, providing scientific evidence for understanding human–primate conflict ecology and developing effective management strategies. Full article
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24 pages, 3329 KB  
Article
Isolation of a Novel Streptomyces sp. TH05 with Potent Cyanocidal Effects on Microcystis aeruginosa
by Xuhan Wang, Siqi Zhu, Shenchen Tao, Shaoyong Zhang, Ruijun Wang and Liqin Zhang
Toxins 2025, 17(7), 354; https://doi.org/10.3390/toxins17070354 - 17 Jul 2025
Viewed by 735
Abstract
In this paper, cultivable actinobacteria were isolated, cultured, and identified from the heavily algal-bloomed waters of Taihu Lake using 16S rRNA gene sequencing. Among the isolates, a single strain exhibiting vigorous cyanocidal activity against Microcystis aeruginosa FACHB-905 was selected for further investigation. The [...] Read more.
In this paper, cultivable actinobacteria were isolated, cultured, and identified from the heavily algal-bloomed waters of Taihu Lake using 16S rRNA gene sequencing. Among the isolates, a single strain exhibiting vigorous cyanocidal activity against Microcystis aeruginosa FACHB-905 was selected for further investigation. The cyanocidal efficacy and underlying mechanisms of this strain, designated TH05, were assessed through using chlorophyll content, cyanobacterial inhibition rate, and cyanobacterial cell morphology measurements. In addition, oxidative stress responses, expression of key functional genes in FACHB-905, and variations in microcystin concentrations were comprehensively evaluated. Cyanobacterial blooms caused by Microcystis aeruginosa pose serious ecological and public health threats due to the release of microcystins (MCs). In this study, we evaluated the cyanocidal activity and mechanism of a novel actinomycete strain, Streptomyces sp. TH05. Optimization experiments revealed that a light–dark cycle of 12 h/12 h, temperature of 25 °C, and pH 7 significantly enhanced cyanocidal efficacy. Under these conditions, TH05 achieved an 84.31% inhibition rate after seven days of co-cultivation with M. aeruginosa. Scanning electron microscopy revealed two distinct cyanocidal modes: direct physical attachment of TH05 mycelia to cyanobacterial cells, causing cell wall disruption, and indirect membrane damage via extracellular bioactive compounds. Biochemical analyses showed increased levels of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) during the first five days, peaking at 2.47-, 2.12-, and 1.91-fold higher than control levels, respectively, indicating elevated oxidative stress. Gene expression analysis using elf-p as a reference showed that TH05 modulated key genes associated with photosynthesis (PsaB, PstD1, PstD2, RbcL), DNA repair and stress response (RecA, FtsH), and microcystin biosynthesis (McyA, McyD). All genes were upregulated except for RbcL, which was downregulated. In parallel, microcystin content peaked at 32.25 ng/L on day 1 and decreased to 16.16 ng/L by day 9, which was significantly lower than that of the control group on day 9 (29.03 ng/L). These findings suggest that strain TH05 exhibits potent and multifaceted cyanocidal activity, underscoring its potential for application in the biological control of cyanobacterial blooms. Full article
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Article
A Quantitative Method for Characterizing of Structures’ Debris Release
by Maiqi Xiang, Martin Morgeneyer, Olivier Aguerre-Chariol, Caroline Lefebvre, Florian Philippe, Laurent Meunier and Christophe Bressot
Eng 2025, 6(7), 157; https://doi.org/10.3390/eng6070157 - 10 Jul 2025
Viewed by 258
Abstract
The characterization of airborne submicrometric composite structures’ debris is a challenge in the field of environmental monitoring and control. The work presented here aims to develop a new quantitative method to measure elemental mass concentrations via particle sampling and Transmission Electron Microscopy—Energy-Dispersive X-ray [...] Read more.
The characterization of airborne submicrometric composite structures’ debris is a challenge in the field of environmental monitoring and control. The work presented here aims to develop a new quantitative method to measure elemental mass concentrations via particle sampling and Transmission Electron Microscopy—Energy-Dispersive X-ray Spectroscopy (TEM-EDS). The principle is to collect airborne particles on a porous TEM grid, then add a certain mass of reference particles, and compare the relative mass percentages of elements from reference and sample particles via EDS. Diverse pairs of airborne particles (RbCl, CsCl, NaCl, SrCl2, Ga(NO3)3, braking particles) were deposited on one TEM grid, and the experimental elemental mass ratios were measured by EDS and compared with the theoretical values. Results show that the quantitative and homogeneous collection of reference particles, such as RbCl, on the TEM grid could be suitable. For all the tested conditions, the absolute deviations between the theoretical elemental mass ratios and the experimental ratios remain lower than 8%. Thus, the mass concentration of Fe from the braking aerosol is calculated as 107 µg/m3. Compared to the cumbersome real-time instrument, this new method for mass characterization appears to be convenient, and requires a short time of aerosol sampling at the workplace. This approach ensures safety and practicability when assessing, e.g., the exposure risk of hazardous materials. Full article
(This article belongs to the Section Materials Engineering)
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