Search Results (741)

The recent rise in whole blood usage for traumatic hemorrhagic shock has renewed interest in the impact of leukocytes on hemostatic function during cold storage. This study investigated whether tissue factor (TF)-bearing extracellular vesicles (EVs) are released from human monocytic cells during cold storage or upon rewarming and whether this process is mechanistically linked to apoptosis. We further examined the contribution of superoxide anion generated by NADPH oxidase (NOX). Methods: THP-1 cells were incubated at 4 °C for up to 24 h with/without test reagents and subsequently rewarmed at 37 °C. Cells were washed by centrifugation before rewarming as required. TF activity in the cell supernatant was quantified, EVs were analyzed by flow cytometry with size-defined gating, and NOX activity normalized to p22^phox was measured by cytochrome c reduction. Results: TF levels and apoptotic cells increased during cold storage. TF release was enhanced 1–2 h after cell lavage following cold exposure, indicating active shedding of TF-bearing EVs rather than passive leakage from damaged membranes. Flow cytometry demonstrated that TF-bearing EVs were distinct from apoptotic vesicles, with a substantial proportion falling within the microvesicle size range. Cold exposure enhanced NOX activity. Both superoxide dismutase (SOD) and catalase inhibited TF release during cold storage; however, only SOD suppressed TF release after cell lavage. Conclusions: TF-bearing EVs are actively shed from human monocytic cells during and after cold storage via a NOX-dependent, superoxide-mediated mechanism. Extracellular SOD suppressed this procoagulant EV release, suggesting a potential strategy to modulate hemostatic alterations associated with cold-stored blood. Full article

Alzheimer’s disease (AD) is characterized by the accumulation of amyloid beta (Aβ) and neurofibrillary tangles in brain tissue; however, AD is multifactorial, and different etiopathogenic mechanisms involve factors that can affect mitochondrial function, which are associated with AD. While high-dose lithium is a well-established mood stabilizer, accumulating evidence suggests that low-dose lithium provides significant neuroprotection by reversing AD pathology, cognitive impairment, and inflammation. Despite these findings, there is limited information on how lithium affects brain energy metabolism. In the current study, we investigated the effect of lithium (0, 0.1, 1, and 10 mM) on mitochondrial function in AD neurons. Neuronal cells were isolated from the hippocampi of embryonic day 14–17 (E15–E17) control (C57BL/6) mice and 3xTg-AD mice. Mitochondrial oxygen consumption rate (OCR), mitochondrial Cytochrome C Oxidase (COX) activity, total ATP activity, and the expression of mitochondrial complex protein involved in oxidative phosphorylation (OXPHOS) were measured in control vs. 3xTg-AD in the presence and absence of lithium treatment. In the present study, lithium treatment significantly increased (p < 0.05) mitochondrial OCR, COX, total ATP, and levels of mitochondrial complex protein subunits (Complex I–V) in 3xTg-AD neurons. However, lithium had no effect on energy metabolism in control neurons. Together, these data indicate that lithium improves mitochondrial function under pathological states. Overall, these results have important implications for the treatment of disorders in which brain energy regulation is compromised, including AD. Particularly, our results highlight a role for lithium in regulating bioenergetics in early-stage AD and suggest that neuronal cells may be a crucial therapeutic target for preventing AD. Full article

The durian seed borer, Mudaria stahlgretschae, is a major economic pest that has significantly impacted durian cultivation in Southeast Asia; however, comprehensive biological and ecological data for this species remain limited. This study employs an integrative taxonomic approach, combining morphological examination with molecular validation of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Phylogenetic analysis (Neighbor-Joining) confirmed that all collected specimens (n = 11) formed a distinct monophyletic clade within the genus Mudaria, showing a genetic identity of 95.75–96.85% with existing GenBank accessions, thereby confirming their identity as M. stahlgretschae. Systematic monitoring using light traps in Uttaradit Province revealed a clear seasonal phenology, with adult flight activity restricted to a five-month period from April to July 2025. Population density peaked in May (55.56%), synchronized with the mid-stages of durian fruit development. Furthermore, chemical profiling of female gland volatiles via GC-MS identified 40 compounds; among these, four putative sex pheromone candidates—1-Hexacosene, (Z)-7-Hexadecenal, 11-Octadecenal, and 2-Hexadecanol—were identified as key constituents based on their consistent detection across all replicates (n = 3), high relative abundance, and absence in male extracts or blank controls. These findings establish a critical foundation for developing synthetic pheromone lures and synchronized monitoring programs, offering a robust framework for the sustainable management of M. stahlgretschae in durian agroecosystems. Full article

Mesenchymal stem cells (MSCs) are central effectors in regenerative medicine, yet their clinical translation is hindered by inconsistent therapeutic outcomes and a lack of standardized light-delivery protocols. This review addresses an underexplored dimension of photobiomodulation (PBM): the divergent, wavelength-dependent signaling programs triggered in MSCs by red/near-infrared (NIR) versus blue light. By integrating biophysical principles of light delivery with a systems biology analysis of protein–protein interaction networks (STRING/GO), we delineate a “Dual Photonic Programming” framework. Red/NIR light (600–1100 nm) targets mitochondrial cytochrome c oxidase, activating a bioenergetic-anabolic program centered on PI3K/Akt/mTOR and Wnt/β-catenin—pathways essential for structural tissue repair. Blue light (400–500 nm) engages cytosolic flavins to drive a secretory-paracrine program that modulates vesicle trafficking and immunomodulatory cargo release. We further examine the dosimetric paradox, demonstrating how culture-environment optics and the Arndt–Schultz biphasic law govern the transition from regenerative stimulation to inhibitory oxidative stress. By tailoring photonic parameters to the MSC’s anatomical origin and metabolic baseline, PBM can serve as a high-fidelity bio-switch for orchestrated tissue repair, providing a mechanistic roadmap for standardized regenerative therapies. Full article

Lake Bacalar is an oligotrophic body of water considered an extreme environment due to its unusual chemistry, with many species yet to be discovered. Mysids were recently collected in this system using light traps. Most previous records of these organisms in the continental Yucatán Peninsula are from closed cenotes (sinkholes or dolines) and caves. We applied integrative taxonomy, considering morphological characters of mature males and females, their geographical distribution, and mitochondrial cytochrome C oxidase subunit I (COI) gene sequences to describe Mayamysis bacalarensis n. gen., n. sp. The morphological analyses included scanning electron microscopy (SEM) and bright-field microscopy observations. We found that M. bacalarensis n. gen., n. sp. has a unique combination of morphological characters within the tribe Gastrosaccini, such as the absence of rostrum, the terminal structure of the third pair of pleopods in males, which are highly developed and have a reduced endopod. Both sexes have a lamellar projection on the fifth abdominal segment and numerous setae (35–40) and spines (22) on the margins of the exopod and endopod of the uropods. In addition, the telson has a posterior cleft margin, bearing a total of 32 spines, the apical and lateral ones being the largest. Furthermore, the genetic sequences of these mysids indicate that the organisms from Bacalar are unique, showing an average of 15% genetic divergence from the closest relative, represented by Chlamydopleon dissimile, and over 20% to more than 50% divergence with other mysids sequenced from Mexico. The geographic range of M. bacalarensis n. gen., n. sp. includes the northern part of Chetumal Bay, a brackish oligotrophic lagoon, where organisms identical to those from Lake Bacalar were collected. This finding indicates a possible permanent connectivity through subterranean water flows. Based on the evidence, we conclude that this is a new genus and species, possibly microendemic to southeastern Quintana Roo, Mexico. Full article

Plant-parasitic reniform nematodes of the genus Rotylenchulus are semi-endoparasites of herbaceous and woody plants occurring in regions with Mediterranean, tropical, and subtropical climates. In the present study, the occurrence of reniform nematodes in the rhizosphere of three olive orchards in Central Italy and six in Sicily (Italy) was investigated. Two Rotylenchulus species were recovered in olive groves in Central Italy, and no Rotylenchulus species were found in Sicily. Using the integrative taxonomy approach, combining morphological, molecular and multivariate morphological analyses, the two species were identified as R. borealis and R. macrodoratus. The D2-D3 sequencing of four individual specimens of Italian R. macrodoratus revealed the occurrence of unique haplotypes differing in nucleotide composition each other. Interestingly, the sequencing of different ITS clones from an individual specimen of the Italian R. borealis showed two ITS paralogs differing in length and nucleotide sequence compared with those of other specimens from the same population and showing higher similarity with those from other populations. Phylogenetic analyses, based on D2-D3 expansion domains of the 28S rRNA gene, ITS, and mitochondrial COI, confirmed the high level of ribosomal variability in both species and the occurrence of new mitochondrial haplotypes for the COI. The present study confirms the occurrence of high variability in Rotylenchulus genus and the existence of variant gene copies in the same specimen that could contribute to the survival of these species in different environments. Full article

This study presents the first comprehensive molecular assessment of the Lepidoptera fauna of Austria based on DNA barcodes (cytochrome c oxidase I, COI; 658 bp Folmer region). The barcode reference library comprises approximately 23,500 sequences, representing 3591 Linnaean species or about 85% of the known national species (ca. 4200 species). Congruence between morphological species identifications under the Linnaean system and barcode data was evaluated using the Barcode Index Number (BIN) system. A total of 244 species could not be unambiguously assigned, showing two to seven BINs that exhibit elevated genetic divergence and may partially represent cryptic diversity. These taxa, together with 40 currently unnamed lineages, require further integrative taxonomic assessment. The distinctiveness of the Austrian Lepidoptera fauna is discussed in the context of endemic genetic diversity. Finally, 17 new faunistic records for Austria are reported. Full article

Background/Objectives: Copper (Cu²⁺) is an essential trace element that participates as a cofactor in key metabolic enzymes such as cytochrome c oxidase and superoxide dismutase. However, excessive copper exposure can be toxic and disturbances in copper homeostasis have been associated with neurodegenerative diseases including Alzheimer’s and Parkinson’s disease. Despite growing evidence linking copper to neuronal dysfunction, the cellular mechanisms by which Cu²⁺ affects neuronal excitability and neurotransmission remain poorly understood. The aim of this study was to investigate the effects of acute Cu²⁺ exposure on ionic currents involved in cellular excitability and neurotransmitter release in bovine chromaffin cells. Methods: Primary cultures of bovine chromaffin cells were used as a neuroendocrine model to study cellular excitability. Voltage-dependent ionic currents were recorded using the whole-cell patch-clamp technique in voltage-clamp configuration. Catecholamine secretion was monitored by amperometry, and cytosolic Ca²⁺ dynamics were measured in fluo-4-loaded cells during depolarization induced by high K⁺ stimulation. Results: Acute Cu²⁺ exposure produced a concentration-dependent enhancement of depolarization-evoked catecholamine release. In parallel, Cu²⁺ inhibited voltage-dependent calcium (ICa), sodium (INa), potassium (IKv), and calcium/voltage-dependent potassium (IKCa-v) currents in a concentration-dependent and partially reversible manner. In addition, Cu²⁺ increased basal cytosolic Ca²⁺ levels while reducing the amplitude of depolarization-evoked Ca²⁺ transients. Conclusions: Acute Cu²⁺ exposure exerts a dual effect in bovine chromaffin cells, inhibiting the ionic currents that support cellular excitability while potentiating catecholamine secretion. This apparent paradox is consistent with a disruption of intracellular Ca²⁺ homeostasis, in which elevated basal cytosolic Ca²⁺ may facilitate exocytosis despite reduced depolarization-evoked Ca²⁺ entry. These findings provide new insight into the mechanisms by which copper may alter neuronal signaling and contribute to neurotoxicity. Full article

The ATP-dependent inhibition of cytochrome c oxidase (CytOx, complex IV of the electron transport chain) is the second mechanism of respiratory control adjusting mitochondrial respiration in order to prevent excessive electron flow and reactive oxygen species (ROS) production. Here, we investigate how tricarboxylic acid (TCA) cycle metabolites and the subsequent complex I or complex II activities influence this regulatory mechanism. Therefore, CytOx activity was assessed by the oxygen consumption rate after cytochrome c (Cyt c) titration to stimulate complex IV activity in isolated rat heart mitochondria (RHM) and permeabilized AC16 cells. Mitochondrial membrane potential (Δψm) and ROS formation were analysed by flow cytometry. Our results show that TCA cycle intermediates differed in their impact on CytOx activity and subsequent ROS formation. NADH-linked substrates such as α-ketoglutarate, glutamate and malate increased respiratory capacity, but preserved ATP-dependent control of CytOx, indicating that elevated electron supply alone does not necessarily abolish ATP sensitivity. In contrast, succinate, which feeds electrons directly into complex II, strongly increased respiration causing the loss of ATP-dependent respiratory control in both model systems. Despite this strong respiratory effect, succinate induced only modest changes in mitochondrial membrane potential in isolated mitochondria, whereas permeabilized cardiomyocytes exhibited reduced polarization accompanied by increased superoxide formation. Together, these findings demonstrate that the effectiveness of ATP-dependent CytOx inhibition is influenced by TCA cycle activity and depends on the site of electron entry into the respiratory chain. Thus, substrate-dependent modulation of respiratory control links metabolite availability to mitochondrial redox regulation in cardiac cells. Full article

Environmental DNA (eDNA) metabarcoding has emerged as a powerful tool for biodiversity monitoring, yet its accuracy is fundamentally constrained by the completeness and taxonomic reliability of reference sequence databases. For the Three Gorges Reservoir (TGR), no integrated multi-marker eDNA reference library exists, hampering standardized fish conservation monitoring under the Yangtze River Ten-Year Fishing Ban. Here, we constructed a comprehensive, multi-marker eDNA reference database for the fish fauna of the TGR, encompassing mitochondrial 12S rRNA, 16S rRNA, and cytochrome c oxidase subunit I (COI) gene sequences from 173 specimens (120 species) collected between 2021 and 2024. After integrating publicly available sequences, the final database comprised 161 species. Then, we quantitatively compared species annotation performance between this local database and public repositories. Results showed that while public databases achieved higher nominal species coverage (94.67%), they exhibited critical deficiencies in annotation accuracy, correctly annotating only 77.97% (12S rRNA), 75.00% (16S rRNA), and 38.14% (COI) of sequences from shared species under controlled conditions. In contrast, the local database exhibited 92.37%, 93.10% and 100% annotation accuracy for the respective markers. Optimal interspecific Kimura 2-parameter (K2P) thresholds for species delimitation were 0.00448 (12S rRNA), 0.00531 (16S rRNA), and 0.00734 (COI). In addition, 15, 0, and 4 species pairs exhibited zero interspecific distance for 12S rRNA, 16S rRNA, and COI, respectively. These limitations reinforce the need for cautious interpretation of eDNA metabarcoding results and the integration of multiple markers or complementary nuclear loci. This study provides preliminary evidence that regionally curated, multi-marker reference libraries could improve taxonomic assignment reliability in eDNA metabarcoding compared to uncurated public repositories, providing a foundational resource for biodiversity conservation. Full article

This work introduces an approach to evolutionary analysis in which information encoded in amino-acid sequences is converted into a specific type of image, termed a genetic image. Genetic images derived from the amino-acid sequences of cytochrome b and cytochrome c oxidase subunit I are shown to be suitable for identifying evolutionary similarities between organisms. Furthermore, artificial neural networks are demonstrated to recognize these genetic images, enabling identification of species evolution. The results indicate the similarity of the genetic images of organisms belonging to species that emerged earlier during Earth’s evolutionary history to the genetic images of organisms belonging to species that emerged later. This finding indicates that genetic images are inherited and undergo gradual modification during evolutionary processes. The phenomenon of inheritance and modification of genetic images suggests that evolution tends to change the already existing functionalities of organisms, which allows for the ordering of organisms belonging to different species from ancient forms, through species that appeared successively during evolution, to those belonging to species that have developed more recently, up to Homo sapiens. Moreover, unlike analyses based on phylogenetic trees, the method presented in this article does not require computing hypothetical taxonomic units to study evolution. Combined with analyses of the inheritance of genetic images, it can support the interpretations of phylogenetic trees and evolutionary research. Full article

Background: Pangolins are critically endangered mammals, and a comprehensive understanding of their genetic diversity is crucial for effective conservation. The mitochondrial genome serves as a vital molecular marker for phylogenetic and population genetic studies. Obtaining genetic material from these elusive animals non-invasively remains a challenge. This study aimed to sequence and characterize the complete mitochondrial genome of Manis javanica and explore the phylogenetic relationships among pangolin species. Methods: The complete mitochondrial genome was sequenced from a saliva-derived sample. Standard procedures for DNA extraction, amplification, and sequencing were employed. The genome was assembled and annotated using bioinformatic tools. Phylogenetic analysis was conducted based on the cytochrome c oxidase subunit I (COXI) gene sequences from nine pangolin species, with the resulting tree constructed using the maximum-likelihood method. Results: The complete mitochondrial genome of M. javanica (GenBank accession: PP110760) is a circular molecule of 16,573 bp, containing 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and a control region. The overall base composition showed a lower GC content (43.83%) than AT content (56.17%). Phylogenetic analysis based on COXI sequences delineated the nine species into three distinct genera: Manis, Phataginus, and Smutsia. Within the genus Manis, Manis pentadactyla was identified as the closest relative to M. javanica. The newly described species Manis mysteria was found to be closer to Manis culionensis and Manis crassicaudata than to other congeners. Furthermore, the analysis indicated that African pangolins diverged earlier than Asian pangolins. Conclusions: This study successfully demonstrates the feasibility of extracting and sequencing the complete mitochondrial genome from saliva samples, providing a valuable non-invasive method for future genetic studies on pangolins. The genomic data and phylogenetic results offer significant molecular insights that will benefit the genetic management and conservation of critically endangered pangolin resources. Full article

Furuncular myiasis is rarely reported in Southeast/Eastern Europe and may be underrecognized or misdiagnosed in non-endemic settings. We described two imported furuncular myiasis cases diagnosed in Romania following travel to Peru and confirmed the etiologic agent by larval morphology and mitochondrial cytochrome c oxidase subunit I (COI) sequencing. We also conducted a narrative review of published case reports/series from Southeast/Eastern Europe (1900–2025) and summarized case characteristics. A previously healthy 31-year-old woman and 32-year-old man presented with painful furuncle-like lesions on the upper back near the shoulder and the posterolateral upper arm, respectively, associated with pruritus and a sensation of movement. Each lesion had a central punctum with intermittent air bubbles. Occlusion of the breathing pore with petroleum jelly facilitated mechanical extraction of one barrel-shaped larva per lesion. Microscopy showed features consistent with second-instar Dermatobia hominis larvae, and COI sequencing demonstrated 97.14–99.33% identity with reference D. hominis sequences. Literature review identified 25 travel-associated cases, with D. hominis involved mostly after travel to Central/South America. These cases highlight the value of travel history and key diagnostic clues for D. hominis myiasis in travelers that may enable timely diagnosis and minimally invasive management. Greater awareness and reporting are needed to better define epidemiology. Full article

The Rhiniinae (Diptera: Calliphoridae), a recently reclassified subfamily of blowflies, comprise approximately 400 species across 30 to 39 genera, which occupy diverse ecological associations, including flower visitation and specialized associations with social insects, yet their phylogenetic relationships remain poorly established due to lack of molecular data. We sequenced and characterized the complete mitochondrial genome of six representative Rhiniinae species, with which the phylogenetic analyses were conducted. The monophyly of Rhiniinae was robustly supported and the internal relationships were clarified. Rhiniini and Cosminini were both recovered as well-supported monophyletic tribes using comprehensive mitogenomic evidence for the first time. In contrast to the purifying selection prevailing in most protein-coding genes, the COII gene showed consistent signatures of positive selection, potentially linked to the functional optimization of cytochrome c oxidase. Overall, this study provides foundational mitogenomic data and a robust phylogenetic framework, offering valuable resources for future research on mitochondrial evolution and systematics within this ecologically intriguing lineage. Full article

To address cryodamage in Culter alburnus sperm, this study evaluated the effects of trehalose supplementation in a conventional cryomedium (D-15 + 10% ethylene glycol). Six experimental groups were established: fresh sperm (G1), a conventional cryomedium (G2), groups supplemented with 10, 100, or 200 mmol/L trehalose (G3–G5), and a control group with extender only (G6). The group with 100 mmol/L trehalose (G4) was associated with improved post-thaw motility parameters (activation rate, movement time, and lifespan) and higher antioxidant (superoxide dismutase and catalase) and energy metabolism (ATPase, succinate dehydrogenase, lactate dehydrogenase) enzyme activities. Ultrastructural damage in G4 included partial plasma membrane rupture and mitochondrial swelling, while G6 exhibited additional damage features including membrane disintegration, mitochondrial disruption, and flagellar fracture. Proteomic analysis revealed that, compared to G1, G4 exhibited higher abundance of proteins (e.g., Histone H2A, cytochrome c oxidase, profilin) involved in structural integrity and energy homeostasis, whereas G6 showed signatures of oxidative stress and metabolic dysfunction (lower abundance of NADH dehydrogenase and higher abundance of calcium-transporting ATPase and glutathione S-transferase). In conclusion, 100 mmol/L trehalose was associated with improved cryopreservation outcomes, and the proteins identified provide a basis for further investigation. This approach offers a framework for refining germplasm conservation strategies in aquaculture. Full article