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With their diverse species, mosquitoes are known to transmit the causal agents of diseases such as malaria, dengue, and yellow fever. Their high adaptability, attraction to humans, and variable adult behaviors make them a significant health concern. The focus on Aedes aegypti is significant for reducing vector-human contacts, monitoring insecticide resistance, and developing innovative vector management strategies. Given the scarcity of studies on Ae. aegypti in the western region of Saudi Arabia, this research is a significant step forward. The study aims to analyze the genetic variations and conduct a phylogenetic study of forty Ae. aegypti samples collected from Taif and Jeddah governorates of Saudi Arabia. The mitochondrial cytochrome c oxidase subunit I (COI) locus was targeted for genetic variance and phylogenetic analysis. Sequences of COI of Ae. aegypti isolates were submitted to the DNA Data Bank of Japan (DDBJ) and National Center for Biotechnology Information (NCBI) Genbank and compared with other global Aedes species isolates. The phylogenetic analysis shows that Ae. aegypti samples from Jeddah have identities ranging from 96.9% to 99.8%, closely related to the Peru (MN299016) and Cambodia (MN299014) isolates. Taif isolates have genetic similarities ranging from 97.5% to 99.8%, closely related to the Germany (KY022526) isolate. Sequence alignment and pairwise comparison show variation among the populations of Ae. aegypti from Taif and Jeddah regions (74.24-98.84%) with a genetic divergence distance of 0.008-0.12. In comparison, ranges slightly change with other Ae. aegypti (79.92-95.96%, 0.008-0.01) as well as Ae. albopictus populations (74.13-83.58%, 0.13-0.20) found in the Genbank database. According to our findings, the present study provides information for a local variation of Ae. aegypti in the western region of Saudi Arabia that could help in disease mapping and risk mitigation, thereby enhancing our ability to manage disease vectors effectively., (Copyright © 2024 by The American Mosquito Control Association, Inc.)

There are hundreds of rare syndromic diseases involving hearing loss, many of which are not targeted for clinical genetic testing. We systematically explored the genetic causes of undiagnosed syndromic hearing loss using a combination of whole exome sequencing (WES) and a phenotype similarity search system called PubCaseFinder. Fifty-five families with syndromic hearing loss of unknown cause were analyzed using WES after prescreening of several deafness genes depending on patient clinical features. Causative genes were identified in 22 families, including both established genes associated with syndromic hearing loss (PTPN11, CHD7, KARS1, OPA1, DLX5, MITF, SOX10, MYO7A, and USH2A) and those associated with nonsyndromic hearing loss (STRC, EYA4, and KCNQ4). Association of a DLX5 variant with incomplete partition type I (IP-I) anomaly of the inner ear was identified in a patient with cleft lip and palate and acetabular dysplasia. The study identified COL1A1, CFAP52, and NSD1 as causative genes through phenotype similarity search or by analogy. ZBTB10 was proposed as a novel candidate gene for syndromic hearing loss with IP-I. A mouse model with homozygous Zbtb10 frameshift variant resulted in embryonic lethality, suggesting the importance of this gene for early embryonic development. Our data highlight a wide spectrum of rare causative genes in patients with syndromic hearing loss, and demonstrate that WES analysis combined with phenotype similarity search is a valuable approach for clinical genetic testing of undiagnosed disease., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics approval: This study was performed in line with the principles of the Declaration of Helsinki. This study was approved by the institutional ethics review board at the National Hospital Organization Tokyo Medical Center (R21-053), RIKEN Center for Integrative Medical Sciences (Yokohama H24-15(3)), and the Tokyo Medical and Dental University (O2015-503). All the experimental procedures for generation and use of Zbtb10-deficient mice were approved by National Hospital Organization Tokyo Medical Center (14-animal-05). Consent to participate: Studies were conducted only after patients or their parents signed informed consent forms to participate. Consent to publish: The participant has consented to the submission of the case report to the journal., (© 2025. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Dyneins are huge motor protein complexes that are essential for cell motility, cell division, and intracellular transport. Dyneins are classified into three major subfamilies, namely cytoplasmic, intraflagellar-transport (IFT), and ciliary dyneins, based on their intracellular localization and functions. Recently, several near-atomic resolution structures have been reported for cytoplasmic/IFT dyneins. In contrast, the structures of ciliary dyneins, as well as their regulatory mechanisms, have yet to be fully elucidated. Here, we isolated a heterodimeric ciliary dynein (IDA-f/I1) from Chlamydomonas reinhardtii, a ciliated green alga, and studied its structure in the presence or absence of ATP by negative-stain electron microscopy and single-particle analysis. Surprisingly, a population of IDA-f adopted a distinctive compact structure, which has been scarcely reported for ciliary dyneins but is very similar to the "phi-particle" structure widely recognized as the autoinhibited/inactivated conformation for cytoplasmic/IFT dyneins. Our results suggest that the inactivation mechanism of dimeric dyneins is conserved in all three dynein subfamilies, regardless of their cellular functions, highlighting the intriguing intrinsic regulatory mechanism that may have been acquired at an early stage in the evolution of dynein motors., (© 2025 Wiley Periodicals LLC.)

Background: In response to iron deficiency and other environmental stressors, cyanobacteria producing siderophores can help in ameliorating plant stress and enhancing growth physiological and biochemical processes. The objective of this work was to screen the potential of Arthrospira platensis, Pseudanabaena limnetica, Nostoc carneum, and Synechococcus mundulus for siderophore production to select the most promising isolate, then to examine the potentiality of the isolated siderophore in promoting Zea mays seedling growth in an iron-limited environment., Results: Data of the screening experiment illustrated that Synechococcus mundulus significantly recorded the maximum highest siderophore production (78 ± 2%) while the minimum production was recorded by Nostoc carneum (24.67 ± 0.58%). Therefore, Synechococcus mundulus was chosen for the beneficiary study and the intended agricultural application. Siderophore-type identification tests proved that Synechococcus mundulus produced hydroxamate-type. The response surface approach was successful in optimizing the conditions of siderophore production in Synechococcus mundulus with actual values for maximum biomass (387.11 mg L - 1 ) and siderophore production (91.84%) higher than the predicted values. The proton nuclear magnetic resonance ( 1 H NMR) analysis data and the Fourier transformer-infrared spectrum analysis (FT-IR) signify the hydroxamate nature of Synechococcus mundulus isolated siderophore. Zea mays seedlings' growth response in the hydroponic system was significantly stimulated in response to supplementation with Synechococcus mundulus siderophore in the absence of iron compared to plants grown without iron and the positive controls. Additionally, the contents of chlorophyll a, chlorophyll b, carotenoids, total carbohydrates, and total protein were all surpassed in siderophore-treated plants, which is expected due to the increased iron content., Conclusions: The results introduced in this study highlighted the significant potential of Synechococcus mundulus-derived siderophore in stimulating Zea mays physicochemical growth parameters and iron uptake. Findings of this study present novel visions of cyanobacteria producing siderophores as an ecofriendly alternative candidate to synthetic iron chelators and their role in plant stress management., Competing Interests: Declarations. Ethics approval and consent to participate: All authors read and approved of the final manuscript and related ethics. Consent for publication: All authors consent to this manuscript for publication after revising this final form. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Norovirus (NoV) infection is a leading cause of gastroenteritis and poses global health threats, with increasing incidence reported in immunocompromised individuals, which is further exacerbated by the globalization of the food industry. Eumelanin has demonstrated its potential in antiviral treatments, but its role in preventing viral infections remains underexplored. Therefore, in this study, we investigated the antiviral properties and potential mechanisms of self-assembled eumelanin nanoparticles (EmNPs) against Tulane virus (TuV), a surrogate with a similar infection mechanism to NoVs. EmNPs exhibited low cytotoxicity and strong antiviral activity in pre-incubated cells. Additionally, EmNPs stimulated the growth and endocytosis of cilia on the cell surface, exposing internal long-nanoparticle chains to interact with the cell membrane while promoting cilia growth and enhancing intercellular connections in cells. EmNPs were then transported to lysosomes via vesicles, leading to a perinuclear lysosome clustering. EmNPs activated several key intracellular signaling pathways, including Toll-like receptor (TLR) and C-type lectin receptor (CLR) pathways, along with activating NF-κB, Rap1, TNF, and Hippo pathways. This regulatory action initiated innate cellular immunity, significantly enhancing the production of type I interferons (IFN-α/β) and promoting the localization of lysosomes to the perinuclear region. Therefore, this study illustrated that EmNPs effectively stimulated immune responses, improved intercellular communication, and facilitated transport mechanisms, thereby bolstering resistance to subsequent viral infections. These findings position EmNPs as promising candidates for the prevention of norovirus infections.

In this study, an alginate-chitosan (AL-CS) nanocomplex decorated with vitamin C coated iron oxide nanoparticles (Fe 3 O 4 -vit C NPs) was investigated as a novel nanoiron fortification agent. The Fe 3 O 4 -vit C NPs decorated on AL-CS nanocomplex underwent comprehensive characterization, including zeta potential, fourier transform infrared spectroscopy, X-ray diffraction, and UV-vis spectroscopy. The transmission electron microscopy (TEM) analysis confirmed the decoration of Fe 3 O 4 -vit C NPs on AL-CS nanocomplex. The dynamic light scattering and thermogravimetric analysis showed enhanced thermal properties of decorated nanocomplex than the undecorated control. Biocompatibility testing on HepG2 cell lines revealed improved compatibility, while intestinal Caco2 cell lines showed approximately 51 % greater bioacessibility than controls. Further, 8 mg of Fe 3 O 4 -vit C NPs decorated AL-CS nanocomplex nanofortified 80 g of A2 goat milk gels (GMGs) which provided 0.072 mg/g of nanoiron without showing significant changes in texture and color compared to the control A2 GMGs. The PCA analysis helped to identify the impact of various factors for the preparation of decorated nanocomplex., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Shweta Rathee reports a relationship with National Institute of Food Technology Entrepreneurship and Management that includes: non-financial support. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

During anuran metamorphosis from herbivorous tadpoles to carnivorous frogs, the gastrointestinal (GI) tract undergoes drastic remodeling, such as the formation of the stomach-intestine boundary and the development of the pyloric sphincter at the posterior end of the stomach. However, the morphogenetic process and molecular mechanisms of how the pyloric sphincter is formed during metamorphosis, instead of during embryogenesis as in amniotes, are largely uninvestigated. Using the African clawed frog Xenopus laevis, we histologically examined the development of the pylorus region from embryonic to froglet stages and performed spatiotemporal gene expression analyses. We found that the pyloric sphincter is formed at a flexure within the pyloric region during metamorphic climax, and that the pyloric and duodenal epithelia, which are morphologically indistinguishable before sphincter formation, become clearly demarcated by the sphincter at the end of metamorphosis. Consistent with these morphological changes, expression domains of a stomach marker barx1 and an intestine marker cdx2 overlapped until late metamorphic climax, but became separated after metamorphosis. Despite the absence of the sphincter before metamorphosis, various genes crucial for sphincter formation in amniotes were already expressed in the pylorus region of Xenopus embryos. RNA-sequencing analysis at pre-metamorphic and metamorphic-climax stages suggest unappreciated roles of genes, such as those for retinoic acid signaling and various transcription factors, in suppressing or promoting sphincter formation. These data provide histological and molecular insights into the heterochrony of the pyloric sphincter formation in amniotes and anurans., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This review paper analyzes recent advancements in bio-polymer coatings for probiotic microencapsulation, with a particular emphasis on chitosan and its synergistic combinations with other materials. Probiotic microencapsulation is essential for protecting probiotics from environmental stresses, enhancing their stability, and ensuring effective delivery to the gut. The review begins with an overview of probiotic microencapsulation, highlighting its significance in safeguarding probiotics through processing, storage, and gastrointestinal transit. Advances in chitosan-based encapsulation are explored, including the integration of chitosan with other bio-polymers such as alginate, gelatin, and pectin, as well as the application of nanotechnology and innovative encapsulation techniques like spray drying and layer-by-layer assembly. Detailed mechanistic insights are integrated, illustrating how chitosan influences gut microbiota by promoting beneficial bacteria and suppressing pathogens, thus enhancing its role as a prebiotic or synbiotic. Furthermore, the review delves into chitosan's immunomodulatory effects, particularly in the context of inflammatory bowel disease (IBD) and autoimmune diseases, describing the immune signaling pathways influenced by chitosan and linking gut microbiota changes to improvements in systemic immunity. Recent clinical trials and human studies assessing the efficacy of chitosan-coated probiotics are presented, alongside a discussion of practical applications and a comparison of in vitro and in vivo findings to highlight real-world relevance. The sustainability of chitosan sources and their environmental impact are addressed, along with the novel concept of chitosan's role in the gut-brain axis. Finally, the review emphasizes future research needs, including the development of personalized probiotic therapies and the exploration of novel bio-polymers and encapsulation techniques., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Breast cancer (BC) has a prevalence rate of 21.8% among Saudi women and ranks as the third leading cause of death in Western nations. Nanotechnology offers innovative methods for targeted BC therapy, and this study explores the use of single-walled carbon nanotubes (SWCNTs) for delivering the senna leaf extract. The study evaluated the effects of increasing dosages of senna leaf extract conjugated to SWCNTs on MCF-7 cells. Cell viability was assessed using the MTT assay, while Giemsa staining revealed morphological changes. Additionally, the comet assay and agarose gel electrophoresis were employed to evaluate the pro-apoptotic potential. The potential of mitochondrial membrane and the production of reactive oxygen species (ROS) were investigated using the JC-1 dye. The results indicated that treated cells exhibited apoptotic characteristics, including elevated ROS levels and decreased mitochondrial membrane potential. In summary, the application of nanotechnology to deliver the senna leaf extract shows promise as a herbal treatment for BC, suggesting a potential breakthrough in combating this widespread and deadly disease., Competing Interests: Conflict of interest: Authors state no conflict of interest., (© 2024 the author(s), published by De Gruyter.)

The Kingdom of Eswatini is a Party to the Convention on Biological Diversity and to the Cartagena Protocol on Biosafety. As Party, Eswatini has domesticated these agreements by passing the Biosafety Act, of 2012 to provide for the safe handling, transfer, and use of living modified organisms (LMOs) in the country. The Act regulates living modified organisms to be used for confined field trials, commercial release, import, export, and transit, and for food, feed, and processing. Guidance is provided for prospective applicants before any application is made to the Competent Authority. This framework also provides for the regulation of emerging technologies such as synthetic biology and genome editing. The regulatory framework for living modified organisms aims to provide an enabling environment for the precautionary use of modern biotechnology and its products in the country in order to safeguard biological diversity and human health.

During avian germ cell formation, primordial germ cells (PGCs) differentiate into prospermatogonia in testicular seminiferous tubules or into oogonia in the ovarian cortex in late-stage embryos. Although estrogenic endocrine-disrupting chemicals (EDCs) have been suggested to affect the differential fate of avian germ cells, there is currently no established method to examine the effects of EDCs on the differentiation potential of germline cells due to large amount of unidentified proteins present in avian germ cells. Regarding reliable molecular probes for the detection of germ cells that differentiated from the PGCs of Japanese quail, the prospermatogonium and oogonium, respectively, integrin beta1 (ITGB1), insulin-like growth factor 2-binding protein 1 (IGF2BP1), and stimulated by retinoic acid 8 (STRA8) were identified as marker proteins by RNA-seq and liquid chromatography tandem mass spectrometry analyses. This study also showed disordered germ cell formation in ovo following the addition of 100 nmol of diethylstilbestrol (DES), o, p'-dichloro-diphenyl-trichloroethane (o,p'-DDT), ethinylestradiol (EE), and bisphenol A. DES directly induced severe damage in germline cells by inhibiting their proliferation and subsequent differentiation into ITGB1-positive germ cells in males independently of disordered gonadal differentiation, while DES and o,p'-DDT decreased the number of female germ cells. In addition, EE toxicity was characterized by a reduction in IGF2BP1-germ cells due to the partial ovarian-like differentiation of male gonads. Furthermore, all EDCs exerted deleterious effects on female ovaries, which restricted differentiation into STRA8-positive oogonia. These results demonstrate that the bioaccumulation of estrogenic EDCs in birds during incubation may reduce male and female fertility., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

A circadian clock is reconstituted in vitro by incubating three proteins, KaiA, KaiB, and KaiC from the non-nitrogen-fixing cyanobacterium Synechococcus elongatus PCC 7942 in the presence of ATP. Leptolyngbya boryana is a filamentous cyanobacterium that grows diazotrophically under microoxic conditions. Among the aforementioned proteins, KaiC is the main clock oscillator belonging to the RecA ATPase superfamily. Genomic studies have revealed the presence of many genes encoding KaiC family ATPases in archaea and bacteria; however, very few have been analyzed in detail. For example, the L. boryana genome encodes two kaiC homologs designated as LbkaiC1 (LBWT_14830) and LbkaiC2 (LBWT_17950). LbKaiC1 is highly similar to KaiC from S. elongatus PCC 7942 compared with LbKaiC2. LbKaiC1 and LbKaiC2 were purified as Strep-tag fusion proteins. LbKaiC1 formed a hexamer and exhibited autophosphorylation, autodephosphorylation, and ATPase activities. Furthermore, it exhibited circadian phosphorylation rhythm in the presence of KaiA and KaiB from S. elongatus PCC 7942, indicating that LbKaiC1 is the central oscillator of the circadian clock in L. boryana. The temporal separation of nitrogen fixation from photosynthesis may be supported by the circadian rhythm generated by LbKaiC1 in L. boryana. LbKaiC2 had low ATPase activity, which depended on temperature, and its autophosphorylation activity was not detected like a circadian oscillator KaiC. Although the function of LbKaiC2 remains unknown, this work will provide comprehensive understanding of KaiC family ATPases., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: GLP-1 receptor agonists, traditionally used for treating type 2 diabetes mellitus and obesity, have demonstrated anti-inflammatory properties. However, their potential neuroprotective effects in neurodegenerative disorders remain unclear., Objective: To evaluate the impact of GLP-1 receptor agonists on the risk of developing various neurodegenerative conditions in obese patients., Methods: This comprehensive retrospective cohort study analyzed data from 5,307,845 obese adult patients across 73 healthcare organizations in 17 countries. Propensity score matching was performed, resulting in 102,935 patients in each cohort. We compared the risk of developing neurodegenerative disorders between obese patients receiving GLP-1 receptor agonist therapy and those who were not., Results: Obese patients treated with GLP-1 receptor agonists showed significantly lower risks of developing Alzheimer's disease (RR = 0.627, 95 %CI = 0.481-0.817), Lewy body dementia (RR = 0.590, 95 %CI = 0.462-0.753), and vascular dementia (RR = 0.438, 95 %CI = 0.327-0.588). The risk reduction for Parkinson's disease was not statistically significant overall (RR = 0.784, 95 %CI = 0.580-1.058) but was significant for semaglutide users (RR = 0.574, 95 %CI = 0.369-0.893). Semaglutide consistently showed the most pronounced protective effects across all disorders. Additionally, a significant reduction in all-cause mortality was observed (HR = 0.525, 95 %CI = 0.493-0.558)., Conclusion: This study provides evidence that the effects of GLP-1 receptor agonists may extend beyond their known metabolic and cardioprotective benefits to include neuroprotection, associated with a decreased risk of developing various neurodegenerative disorders. These findings suggest the potential for expanding the therapeutic applications of GLP-1 receptor agonists to improve neurocognitive outcomes. Further research is warranted to elucidate the mechanisms underlying these neuroprotective effects and to explore their clinical applications in neurodegenerative disease prevention and treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

A probiotic is a live bacterium that, when given in sufficient proportions, helps to improve the host's gut health. Kimchi and pearl millet, two traditional foods, were used to isolate probiotic bacterial strains. This study's primary goals were to isolate, identify and analyse the microorganisms for potential probiotic traits, tolerance to gastrointestinal environments, and antimicrobial activity, and produce probiotic capsule. The present investigation resulted with identification of two probiotic strains (KAC1 and PAC1) from conventional foods, such as kimchi and pearl millet porridge. The isolated probiotics were identified as Enterobacteriaceae family by 16S rRNA sequencing and are deposited in GenBank (NCBI), accession numbers OQ629827 (KAC1) and OQ629828 (PAC1), respectively. These strains exhibited the characteristics of possible probiotic traits, such as the ability to tolerate simulated gastric juice, inhibits the growth of pathogenic bacteria, auto-aggregation, co-aggregation, and hydrophobicity. Furthermore, spectroscopic analysis divulges some critical findings which corroborate the results obtained. Finally, capsules containing freeze-dried  probiotics was successfully produced., Competing Interests: Declarations. Conflict of Interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Prostate cancer is a prevalent and highly heterogeneous malignancy that affects men globally. Despite the availability of various treatment targets, Cytochrome P450 (CYP) enzymes have gained significant attention due to their crucial role in metabolizing both endogenous and exogenous compounds. This study explores Diosmetin as a potential CYP antagonist for treating prostate cancer. To evaluate Diosmetin's potential as a CYP antagonist, we employed a comprehensive in silico approach. Molecular docking was conducted using the Glide software to assess the binding affinity of Diosmetin with CYP enzymes, specifically CYP17A1 and CYP19A1, which are associated with prostate cancer. The druglike properties of Diosmetin were evaluated, focusing on its pharmacokinetic attributes. Additionally, Diosmetin's ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) characteristics were analyzed to determine its suitability as a therapeutic agent. Molecular dynamics (MD) simulations were performed using Desmond to assess the stability and persistence of Diosmetin binding with the CYP enzymes over a 200 ns simulation period. Molecular docking studies revealed robust binding affinities between Diosmetin and CYP17A1 (- 11.261 kcal/mol) and CYP19A1 (- 11.145 kcal/mol). Diosmetin demonstrated favorable pharmacokinetic properties and advantageous ADMET characteristics, including high bioavailability, good dispersion, and favorable metabolism. MD simulations indicated persistent binding interactions between Diosmetin and the CYP enzymes throughout the 200 ns simulation, reinforcing the reliability of these interactions. Pharmacoinformatics investigations provide valuable insights into the potential of Diosmetin as a promising lead compound for the development of novel drug candidates against prostate cancer. The strong binding affinity and favorable pharmacokinetic and ADMET profiles suggest that Diosmetin could be an effective CYP antagonist and warrants further investigation as a potential therapeutic agent for prostate cancer., Competing Interests: Declarations. Consent for publication: The authors declare no conflicts of interest. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Cyclophosphamide (CP) is widely used in chemotherapy to treat various types of cancer. However, it is toxic to the liver and other organs. Rosmarinic acid (RA) possesses anti-inflammatory, antioxidant, and cytoprotective properties. This study investigated the protective effects of RA against CP-induced liver injury in mice. Mice were treated with RA (25, 50, and 100 mg/kg) for 15 days and followed by a single injection of CP on day 16th. CP injection resulted in an elevation in serum AST, ALT, and ALP, along with multiple histopathological alterations in the liver. CP also induced increased levels of MDA and NO, associated with declined GSH, SOD and CAT. RA pretreatment prevented liver injury, alleviated the enhanced levels of MDA and NO, and restored antioxidants defenses, hence avoiding the oxidative injury in the liver. Moreover, RA pretreatment attenuated NF-κB p65 and proinflammatory cytokines levels. Liver of CP-injected mice also showed a decrease in Bcl2, accompanied with elevated BAX and caspase-3 expression, an effect that RA pretreatment alleviated. In addition, pretreatment of CP-administrated mice with RA restored the Nrf2 expression in the liver. Taken together, this study suggests a potential application value of RA in preventing CP hepatotoxicity and sheds light on the possible mechanism., Competing Interests: Declarations. Competing interest: The author declares no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Silver nanoparticles (AgNPs) are spherical particles with a number of specific and unique physical (such as surface plasmon resonance, high electrical conductivity and thermal stability) as well as chemical (including antimicrobial activity, catalytic efficiency and the ability to form conjugates with biomolecules) properties. These properties allow AgNPs to exhibit desired interactions with the biological system and make them prospective candidates for use in antibacterial and anticancer activities. AgNPs have a quenching capacity, which produces reactive oxygen species and disrupts cellular processes (such as reducing the function of the mitochondria, damaging the cell membrane, inhibiting DNA replication and altering protein synthesis). In addition, sponge extracts contain biologically active substances with therapeutic effects. Therefore, the concurrent use of these agents may present a potential for the development of novel antitumor and antimicrobial drugs. The present study investigated the cytotoxic effects of AgNPs combined with the extracts from sponge species, Stylissa carteri or Amphimedon chloros , against various cancer cell lines and pathogenic bacterial strains. The present study was novel as it provided a further understanding of the cytotoxicity and underlying mechanisms of AgNPs. Alterations in the properties, such as size, charge and polydispersity index, of the AgNPs were demonstrated after lyophilization. Scanning electron microscopy revealed submicron-sized particles. The cytotoxic potential of AgNPs across various cancer cell lines such as lung, colorectal, breast and pancreatic cancer cell lines, was demonstrated, especially when the AgNPs were combined with sponge extracts, which suggested a synergistic effect. Analysis using liquid chromatography-mass spectrometry revealed key chemical components in the extracts, and molecular docking simulations indicated potential inhibition interactions between a number of the extract components and the epidermal growth factor receptor and tyrosine kinase receptor A. Synergistic antibacterial effects against several bacterial species such as Staphylococcus xylosus , Klebsiella oxytoca , Enterobacter aerogenes , Micrococcus spp. and Escherichia coli , were observed when AgNPs were combined with sponge ethyl acetate extracts. The results of the present study suggested a potential therapeutic application of marine-derived compounds and nanotechnology in combating cancer and bacterial infections. Future research should further elucidate the mechanistic pathways and investigate the in vivo therapeutic efficacy., Competing Interests: The authors declare that they have no competing interests., (Copyright: © 2025 Alqaraleh et al.)

Study Objectives: Astrocytes change their intracellular calcium (Ca 2+ ) concentration during sleep/wakefulness states in mice. Furthermore, the Ca 2+ dynamics in astrocytes vary depending on the brain region. However, it remains unclear whether alterations in astrocyte activity can affect sleep-wake states and cortical oscillations in a brain region-dependent manner., Methods: Astrocyte activity was artificially manipulated in mice using chemogenetics. Astrocytes in the hippocampus and pons, which are 2 brain regions previously classified into different clusters based on their Ca 2+ dynamics during sleep-wakefulness, were focused on to compare whether there are differences in the effects of astrocytes from different brain regions., Results: The chemogenetic activation of astrocytes in the hippocampus significantly decreased the total time of wakefulness and increased the total time of sleep. This had little effect on cortical oscillations in all sleep-wakefulness states. On the other hand, the activation of astrocytes in the pons substantially suppressed rapid eye movement (REM) sleep in association with a decreased number of REM episodes, indicating strong inhibition of REM onset. Regarding cortical oscillations, the delta wave component during non-REM sleep was significantly enhanced., Conclusions: These results suggest that astrocytes modulate sleep-wakefulness states and cortical oscillations. Furthermore, the role of astrocytes in sleep-wakefulness states appears to vary among brain regions., (© The Author(s) 2024. Published by Oxford University Press on behalf of Sleep Research Society.)

The jabuticaba tree leaf, considered an agro-food by-product or waste, was evaluated as a potential unconventional ingredient of bioactive compounds. Its hydroalcoholic extracts were assessed for the phenolic chemical profile by LC-QQQ-MS and antioxidant capacity pre- and post-gastrointestinal digestion (i.e., bioaccessibility), along with determining cytotoxicity effects on cell culture, antihaemolytic and cellular antioxidant activity. A total of 9 phenolic compounds, including ellagic acid and flavonoids (hesperidin) were identified in both crude and digested samples. The bioaccessibility of these compounds was increased after digestion process, which was reflected in the erythrocytes' antioxidant activities, reducing the haemolysis, lipid peroxidation and reactive oxygen species (ROS) generation by more than 50 %. Furthermore, the in vitro digestion diminished the cytotoxicity effect, increasing the cell viability by 71 %. These findings underscore the potential of jabuticaba tree leaf extract as a rich source of antioxidant phenolics, emphasizing their potential and promising applications as a new functional ingredient., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Objective: Herein, we investigate the potential analgesic effect of a newly synthesized chalcone-derived apocynin in a neurogenic pain model., Methods: Molecular docking was used to foretell the apocynin binding features and dynamics with the TRPV1 channel, and the activity was tested in vitro, using transfected HEK 293T cells with the rat TRPV1 receptor. The analgesic effect of apocynin was investigated using a capsaicin-induced pain model. The expression of TRPV1, TRPA1, TRPM8, and MAPKs was assessed by electrophoresis, and immunosorbent assays were performed to quantify the neurotransmitters Substance P, Glutamate, and CGRP. A survival assay using Galleria mellonella was carried out to determine the toxicity., Results: We observed that apocynin exhibits greater thermodynamic stability. Upon apocynin ligand binding, it changes the electrostatic potential for a predominantly electronegative state in the interior and neutral in its external vanilloid pocket. Treatment of apocynin induces antinociceptive effects against the noxious challenge of capsaicin. Histologically, apocynin decreased the number of TRPV1 + immunopositive cells. Electrophoresis showed reduced phosphorylation of p44/42 (ERK1/2) and decreased protein levels of substance P, and CGRP. In the survival assay, apocynin showed low toxicity., Conclusions: In conclusion, we provide proof-of-principles that the newly synthesized apocynin compound effectively prevented nociception in a neurogenic model of orofacial pain., Competing Interests: Declaration of competing interest All authors declare that they have no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

N-acetylneuraminic acid is an active ingredient in tonic foods and an important additive in foods and biopharmaceuticals. To address the limitations of existing methods of N-acetylneuraminic acid quantification, we developed an immunoassay based on antibodies induced in hens using artificial antigen, showing high sensitivity and specificity with no cross-reactivity with eight N-acetylneuraminic acid analogues. An IgY-based indirect competitive enzyme-linked immunosorbent assay showed a detection range of 1.14 to 70.08 ng/mL and a limit of detection of 0.57 ng/mL. In spiked samples, recoveries by the indirect competitive enzyme-linked immunosorbent assay ranged from 74.05% to 110.87% compared with HPLC (73.01% to 108.8%). Consistency between the indirect competitive enzyme-linked immunosorbent assay and HPLC was satisfactory (R 2  = 0.9736), demonstrating this established immunoassay as a rapid and reliable approach for N-acetylneuraminic acid analysis. The assay described in this study provides an important method for the screening of N-acetylneuraminic acid in biological samples and foodstuffs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Chitosan, a biopolymer derived from the deacetylation of chitin found in crustacean shells and certain fungi, has attracted considerable attention for its promising health benefits, particularly in gut microbiota maintenance and immune system modulation. This review critically examines chitosan's multifaceted role in supporting gut health and enhancing immunity, beginning with a comprehensive overview of its sources, chemical structure, and its dual function as a dietary supplement and biomaterial. Chitosan's prebiotic effects are highlighted, with a focus on its ability to selectively stimulate beneficial gut bacteria, such as Bifidobacteria and Lactobacillus, while enhancing gut barrier integrity and inhibiting the growth of pathogenic microorganisms. The review delves deeply into chitosan's immunomodulatory mechanisms, including its impact on antigen-presenting cells, cytokine profiles, and systemic immune responses. A detailed comparative analysis assesses chitosan's efficacy relative to other prebiotics and immunomodulatory agents, examining challenges related to bioavailability and metabolic activity. Beyond its role in gut health, this review explores chitosan's potential as a dual-action agent that not only supports gut microbiota but also fortifies immune resilience. It introduces emerging research on novel chitosan derivatives, such as chitooligosaccharides, and evaluates their enhanced bioactivity for functional food applications. Special attention is given to sustainability, with an exploration of alternative, plant-based sources of chitosan and their implications for both health and environmental stewardship. Also, the review identifies new research avenues, such as the growing interest in chitosan's role in the gut-brain axis and its potential mental health benefits through microbial interactions. By addressing these innovative areas, the review aims to shift the focus from basic health effects to chitosan's broader impact on public health. The findings encourage further exploration, particularly through human trials, and emphasize chitosan's untapped potential in revolutionizing health and disease management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Thermal and chemical ablation techniques may consolidate recurrent metastatic cervical lymph nodes as alternatives to repeat neck dissection in thyroid cancer patients. This meta-analysis aims to compare the efficacy and safety across modalities., Methods: Four databases were searched for studies on radiofrequency (RFA), microwave (MWA), laser (LA), and ethanol ablation (EA) treating metastatic cervical nodes from thyroid cancer. The outcomes analyzed included treatment response, oncologic control, and complications. Random effects meta-analytical pooling was conducted., Results: There were 25 studies (n = 1061 nodes) examining the four ablation methods. Patients showed comparable baseline characteristics and initial lymph node sizes ranging from 0.96 to 1.28 cm. All modalities achieved substantial node volume reduction (88.4 %) and disappearance (62.8 %), with significant biochemical decline (from 6.01 to 1.13 ng/ml, p = 0.18 between groups). MWA showed the highest volume reduction (99.4 %) and disappearance rate (87.6 %) versus slower efficacy of RFA (93.0 %, 72.1 %), LA (77.9 %, 62.5 %), and EA (81.8 %, 58.4 %). New malignancy/metastases risks ranged from 0.03 % to 1.3 % without between-group differences (p = 0.52). Major complications were absent; transient voice changes (0.05%-10.6 %) and neck pain (0.0%-5.9 %) were the main overall complaints. However, overall complication rates significantly varied by modality (1.1%-10.6 %; p = 0.003)., Conclusions: Thermal and chemical ablation is effective in controlling the metastatic disease burden in patients with thyroid cancer, offering a potentially less morbid and non-surgical alternative to re-operation. Additional prospective data could confirm the long-term equivalent of revision neck dissection and stratify patients based on concomitant Hashimoto's and genomic mutations. Clarifying optimal patient selection and standardizing prognostic indexing could further enhance utilization., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Eyespot patterns have evolved in many prey species. These patterns were traditionally explained by the eye mimicry hypothesis, which proposes that eyespots resembling vertebrate eyes function as predator avoidance. However, it is possible that eyespots do not mimic eyes: according to the conspicuousness hypothesis, eyespots are just one form of vivid signals where only conspicuousness matters. They might work simply through neophobia or unfamiliarity, without necessarily implying aposematism or the unprofitability to potential predators. To test these hypotheses and explore factors influencing predators' responses, we conducted a meta-analysis with 33 empirical papers that focused on bird responses to both real lepidopterans and artificial targets with conspicuous patterns (i.e. eyespots and non-eyespots). Supporting the latter hypothesis, the results showed no clear difference in predator avoidance efficacy between eyespots and non-eyespots. When comparing geometric pattern characteristics, bigger pattern sizes and smaller numbers of patterns were more effective in preventing avian predation. This finding indicates that single concentric patterns have stronger deterring effects than paired ones. Taken together, our study supports the conspicuousness hypothesis more than the eye mimicry hypothesis. Due to the number and species coverage of published studies so far, the generalisability of our conclusion may be limited. The findings highlight that pattern conspicuousness is key to eliciting avian avoidance responses, shedding a different light on this classic example of signal evolution., Competing Interests: AM, ML, PP, YY, MS, SN No competing interests declared, (© 2024, Mizuno et al.)

Scientists have shown great interest in traditional plant extracts, particularly Lepidium sativum (LS), Origanum majorana (OM), Ferula hermonis (FH), and Eruca sativa (ES), which are frequ ently used to improve health. Recently, attention has been directed toward their influence on spermatogenesis and male fertility. Hence, the objective of this study was to explore their impact on male rats' fertility. Antioxidant activity and total phenolic content (TPC) were determined, along with the identification and quantification of phenolic compounds. Oral administration of aqueous extracts was performed individually or as a mixture (MIX) at a dose of 100 mg/kg in 28 male Sprague Dawley rats over a 60-day period. Organ weight, sex hormone concentrations, sperm parameters, oxidative stress markers, histological and morphometric analysis, and protein expression levels were investigated. OM and MIX showed the highest TPC and antioxidant activities, and MIX possessed the highest polyphenolic constituents. Elevated serum testosterone, epididymal sperm concentration, testes glutathione levels, and histomorphometric parameters were manifested in all groups, especially in MIX. MIX group also displayed elevated levels of vimentin, protein kinase B, and mTOR expression in the testes, complemented by declined expression of Phosphatase and Tensin Homolog (PTEN). In conclusion, these findings propose that these extracts, especially MIX followed by OM, enhance fertility by stimulating spermatogenesis.

The present study has evaluated different soybean genotypes to understand the salt and drought tolerance mechanisms based on physiological traits (photosynthesis, stomatal conductance, chlorophyll, and cell membrane stability), antioxidant enzymes (superoxide dismutase, catalase, and peroxidase), reactive oxygen species (H 2 O 2 and O 2 •- ), osmolytes (glycine betaine, proline, and Na + ), plant water relations (relative water content, water potential, and solute potential) and expression of related genes ( + ), plant water relations (relative water content, water potential, and solute potential) and expression of related genes ( GmCAT1 , GmPOD1 , GmSOD , GmP5CS , GmNHX1 , GmAKT1 , GmDREB1 ). The experiment was conducted in a two-factorial arrangement using randomized complete block design (RCBD) with genotypes as one factor and salt, drought, and control treatments as the other factor. All physiological traits, relative water content, and water potential decreased significantly in all soybean genotypes due to individual and combined treatments of drought and salt stress, with significantly less decrease in soybean genotypes G4620RX, DM45X61, and NARC-21. Besides that, the activity of antioxidant enzymes, production of ROS, accumulation of osmolytes, solute potential, and Na GmARF1 ). The experiment was conducted in a two-factorial arrangement using randomized complete block design (RCBD) with genotypes as one factor and salt, drought, and control treatments as the other factor. All physiological traits, relative water content, and water potential decreased significantly in all soybean genotypes due to individual and combined treatments of drought and salt stress, with significantly less decrease in soybean genotypes G4620RX, DM45X61, and NARC-21. Besides that, the activity of antioxidant enzymes, production of ROS, accumulation of osmolytes, solute potential, and Na + /K + ratio were increased significantly in all soybean genotypes under salt and water deficit conditions. As a whole, the soybean genotypes G4620RX, DM45X61, and NARC-21 showed the maximum enzymatic activity with less increase in ROS and Na + in addition to a high accumulation of osmolytes and an increase in solute potential. Correspondingly, the genotypes exhibiting high physiological and biochemical tolerance to drought and salt stresses showed the high expression of genes imparting the stress tolerance. Moreover, correlation, heatmap, and principal component analysis further confirmed the varying physiological and biochemical responses of all soybean genotypes under individual and combined applications of drought and salinity stresses. Overall, the present study confirmed that plants opt for the integrated physiological, biochemical, and genetic approaches to counteract the harmful effects of environmental stresses.+ in addition to a high accumulation of osmolytes and an increase in solute potential. Correspondingly, the genotypes exhibiting high physiological and biochemical tolerance to drought and salt stresses showed the high expression of genes imparting the stress tolerance. Moreover, correlation, heatmap, and principal component analysis further confirmed the varying physiological and biochemical responses of all soybean genotypes under individual and combined applications of drought and salinity stresses. Overall, the present study confirmed that plants opt for the integrated physiological, biochemical, and genetic approaches to counteract the harmful effects of environmental stresses., Competing Interests: The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Alzahrani.)

In Lepidoptera (butterflies and moths), the genomic region around the gene cortex is a "hotspot" locus, repeatedly implicated in generating intraspecific melanic wing color polymorphisms across 100 million years of evolution. However, the identity of the effector gene regulating melanic wing color within this locus remains unknown. We show that none of the four candidate protein-coding genes within this locus, including cortex , serve as major effectors. Instead, a microRNA (miRNA), mir-193 , serves as the major effector across three deeply diverged lineages of butterflies, and its role is conserved in Drosophila . In Lepidoptera, mir-193 is derived from a gigantic primary long noncoding RNA, ivory , and it functions by directly repressing multiple pigmentation genes. We show that a miRNA can drive repeated instances of adaptive evolution in animals.

The complex link between COVID-19 and immunometabolic diseases demonstrates the important interaction between metabolic dysfunction and immunological response during viral infections. Severe COVID-19, defined by a hyperinflammatory state, is greatly impacted by underlying chronic illnesses aggravating the cytokine storm caused by increased levels of Pro-inflammatory cytokines. Metabolic reprogramming, including increased glycolysis and altered mitochondrial function, promotes viral replication and stimulates inflammatory cytokine production, contributing to illness severity. Mitochondrial metabolism abnormalities, strongly linked to various systemic illnesses, worsen metabolic dysfunction during and after the pandemic, increasing cardiovascular consequences. Long COVID-19, defined by chronic inflammation and immune dysregulation, poses continuous problems, highlighting the need for comprehensive therapy solutions that address both immunological and metabolic aspects. Understanding these relationships shows promise for effectively managing COVID-19 and its long-term repercussions, which is the focus of this review paper., (© 2024 Wiley Periodicals LLC.)

Introduction: In a difficult environment like surgery with continuous exposure to stress, surgeon's health and well-being is affected. The heart rate variability (HRV) is a valid stress indicator. This study aims to provide descriptive data regarding stress during urological intervention using HRV., Methods: This prospective, unicentric study, included urological interventions from June to December 2023. Surgeons were surveyed on general characteristic, and surgical skills level. Beat-by-beat heart rate (HR) recorded using POLAR A300® chest belt. HRV were analyzed in time (mean HR and the root mean square of successive differences [RMSSD]) and frequency (Fast Fourier transformation spectrum high frequency [FFT spectrum HF]) domain. Their baseline collected outside the operating room, 3 events during intervention examined: incision, closure, and adverse events. Visual stress and difficulty scale during intervention were completed. The differences between baseline and events were calculated., Results: A total of 171 urological interventions were included. Senior surgeons performed 54 (31.58%) and juniors 117 (68.42%). During interventions, seniors had significantly higher median difference of HR (incision: -11.92 [12.74] bpm versus -2.27 [10.99] bpm for juniors, P≤0.0001; closure: -10.73 [13.71] bpm, versus -4.18 [12.30] bpm for juniors, P=0.0005), lower HF (incision: 0.01 [0.07] hz, versus -0.006 [0.03] hz for juniors P≤0.0001; closure: 0.02 [0.06] hz, versus 0.00 [0.02] hz for juniors, P≤0.0001), and lower RMSSD (incision: -3.34 [13.09] ms for seniors versus -7.63 [10.57] ms for juniors, P=0.0098). No significant difference in RMSSD during closure (P=0.2049)., Conclusion: Study demonstrated the possibility to assess HRV in standard medical practice, senior surgeons experienced more physiological changes during interventions., Competing Interests: Disclosure of interest The authors declare that they have no competing interest., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Speckle-type POZ (SPOP) is described as an essential tumor suppressor factor in gastric cancer, colorectal cancer, and prostate cancer (PCa). SPOP gene mutations were reported in primary human PCa. Isocitrate dehydrogenase-1 (IDH1) oncogene mutations were detected in gliomas, acute myeloid leukemia, some benign and malignant cartilaginous tumors, and only 1% of PCa. This study aimed to investigate the prevalence of mutations of SPOP and IDH1 genes in PCa in the Jordanian population. One hundred formalin-fixed paraffin-embedded tissue samples were collected from patients diagnosed with prostate adenocarcinoma. The obtained specimens were subjected to genomic DNA extraction, PCR amplification, and direct sequencing of exons 4, 5, 6, and 7 of the SPOP gene and exon 6 of the IDH1 gene. SPOP gene mutations were found in 17% of PCa cases, while no mutation was detected in the screened exon 6 of the IDH1 gene. Clinicopathological data demonstrated a strong correlation between prostate-specific antigen (PSA) levels and both Gleason score (GS) and the International Society of Urological Pathology (ISUP) grade group (GG). There was no significant correlation between PSA levels and age (p = 0.816) nor there were significant associations for SPOP mutational status with age (p = 0.659), PSA levels (p = 0.395), GS (p = 0.259), and ISUP GG (p = 0.424) in the tested population. The study found a strong correlation between PSA levels and both GS and ISUP GG. It also identified a high frequency (17%) of SPOP gene mutations in Jordanian Arab PCa patients, mainly in exon 7. No IDH1 mutations were detected in exon 6., Competing Interests: Declarations. Consent for Publication: Not applicable. Competing interest: The authors declare no competing interests. Ethical Approval and Consent to Participate: The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Review Board of King Abdullah University Hospital, Jordan University of Science and Technology (IRB Decision reference number: 68/124/2019. Date: June 27, 2019). Patient consent was waived due to the study design being a retrospective study involving laboratory archives and hospital electronic records, and it did not involve human participants. Human Ethics: Patient consent was waived due to the study design being a retrospective study involving laboratory archives and hospital electronic records, and it did not involve human participants, (© 2024. The Author(s).)

The human body contains trillions of cells, classified into specific cell types, with diverse morphologies and functions. In addition, cells of the same type can assume different states within an individual's body during their lifetime. Understanding the complexities of the proteome in the context of a human organism and its many potential states is a necessary requirement to understanding human biology, but these complexities can neither be predicted from the genome, nor have they been systematically measurable with available technologies. Recent advances in proteomic technology and computational sciences now provide opportunities to investigate the intricate biology of the human body at unprecedented resolution and scale. Here we introduce a big-science endeavour called π-HuB (proteomic navigator of the human body). The aim of the π-HuB project is to (1) generate and harness multimodality proteomic datasets to enhance our understanding of human biology; (2) facilitate disease risk assessment and diagnosis; (3) uncover new drug targets; (4) optimize appropriate therapeutic strategies; and (5) enable intelligent healthcare, thereby ushering in a new era of proteomics-driven phronesis medicine. This ambitious mission will be implemented by an international collaborative force of multidisciplinary research teams worldwide across academic, industrial and government sectors., Competing Interests: Competing interests:  R.A. holds shares of Biognosys AG, which operates in the field covered by the article. D.F. is co-founder of MedBiome Inc., a precision nutrition company. K.G. is a shareholder of CYBO, LucasLand, and FlyWorks. T.G. is the founder of Westlake Omics Inc. M.M. is an indirect investor in EvoSep. R.T. is a founder of BayOmics. The other authors declare no competing interests., (© 2024. Springer Nature Limited.)

We report an entoproct epibiotic on the surface of a sea spider (Pycnogonida). The pycnogonid was identified as Nymphon sp. (Nymphonidae). The entoproct was colonial, with three zooids, and was identified both morphologically and by a molecular phylogeny as Barentsia sp. (Barentsiidae). The largest zooid had eight tentacles and was about 0.7 mm long, smaller than for most colonial entoproct species. We determined partial sequences for the 18S rRNA and 28S rRNA genes from the entoproct. In an 18S-based maximum likelihood tree (1507 characters), the entoproct was the sister taxon to Barentsia gracilis .

Cutaneous leishmaniasis (CL) is a widespread disease affecting both humans and animals globally. Currently, common treatments (e.g., glucantime (GC) for CL treatment have many side effects that limit their use. The current experimental study aims to assess the in vitro, in vivo, and potential mechanisms of action of Rhanterium epapposum essential oil (REE) and its main compounds β-Myrcene (MC), camphene (CP), and limonene (LN) alone and in combination against Leishmania major. In vitro effects of REE and its main compounds were evaluated on amastigote forms, infection in macrophages cells stimulation of nitric oxide (NO), and stimulation of the cellular immunity in macrophages. In vivo efficacy of REE and its main constituents was also assessed in mice with CL through evaluating parasite burden, oxidative stress and proinflammatory-related genes. A concentration-dependent reduction in the average number of amastigotes was observed, with statistical significance (p < 0.001); whereas the results revealed synergistic effects when REE, MC and LN were combined with GC. REE and main compounds mainly in combination elicited a dose-dependent elevation in NO production and the expression levels of inducible nitric oxide synthase (iNOS), interferon gamma (IFN-γ), and tumor necrosis factor (TNF-α) genes in macrophages. Notably, mice treated with a combination of REE, MC, and GC showed the complete recovery of CL lesions after 28 days of treatment and resulted in a reduction of tissue malondialdehyde levels and a significant increase (p < 0.001) in the gene expression levels of the antioxidant enzymes. Topical treating CL-infected mice with REE and its main compounds alone particularly in conjunction with GC, significantly increased (p < 0.001) the expression levels of IFN-γ and interleukin (IL-12), while also causing a notable reduction in IL-4 expression. The findings of the current experimental research revealed the high in vitro and in vivo antileishmanial efficacy of REE and its main compounds MC, CP, and LN mainly in combination with GC; which indicated the high synergic effects of these compounds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)