bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular Identification of Borreliella Species in Ixodes hexagonus Ticks Infesting Hedgehogs (Erinaceus europaeus and E. roumanicus) in North-Western Poland
Authors: Valentyna Kolomiiets , Beata Wodecka
Published: December 2024

The western European hedgehog (Erinaceus europaeus) and the northern white-breasted hedgehog (E. roumanicus) are natural hosts of the tick Ixodes hexagonus, the vector of tick-borne pathogens such as the Borreliella bacteria responsible for Lyme disease. The aim of this study was to identify these pathogens in ticks collected from hedgehogs in northwestern Poland and to assess their genetic diversity by molecular analysis of the detected pathogens based on the flaB gene and the mag-trnI intergenic spacer. Among 101 hedgehogs examined, 737 ticks were found on 56 (55.45%) individuals, including 501 females of I. hexagonus. Borreliella spirochete infection was confirmed in 9 females of I. hexagonus (1.8%) obtained from 4 (3.96%) hedgehogs, detecting Borreliella (Bl.) afzelii (8/89%) and Bl. spielmanii (1/11%). Phylogenetic analysis based on the flaB gene and the mag-trnI intergenic spacer showed a lack of diversity in Bl. afzelii detected in I. hexagonus ticks collected from hedgehogs as well as little diversity against reference strains detected in small mammals and ticks collected from them. The results confirm that hedgehogs play an important role in the circulation of the detected spirochete species, at least as hosts of I. hexagonus ticks infected with them, indicating their potential to spread Borreliella spirochetes.
Unraveling the Mysteries of Sleep: Exploring Phylogenomic Sleep Signals in the Recently Characterized Archaeal Phylum Lokiarchaeota near Loki’s Castle
Authors: Pandi-Perumal Seithikurippu R. , Saravanan Konda Mani , Paul Sayan , Spence David Warren , Chidambaram Saravana Babu
Published: December 2024

Sleep is a universally conserved behavior whose origin and evolutionary purpose are uncertain. Using phylogenomics, this article investigates the evolutionary foundations of sleep from a never before used perspective. More specifically, it identifies orthologs of human sleep-related genes in the Lokiarchaeota of the Asgard superphylum and examines their functional role. Our findings indicate that a conserved suite of genes associated with energy metabolism and cellular repair is involved, thus suggesting that sleep plays a primordial role in cellular maintenance. The data cited lend credence to the idea that sleep improves organismal fitness across evolutionary time by acting as a restorative process. Notably, our approach demonstrates that phylogenomics is more useful than standard phylogenetics for clarifying common evolutionary traits. By offering insight into the evolutionary history of sleep and putting forth a novel model framework for sleep research across taxa, these findings contribute to our growing understanding of the molecular foundation of sleep. This study lays the groundwork for further investigations into the importance of sleep in various organisms. Such investigations could have consequences for improving human health and more generally could provide a deeper comprehension of the fundamental processes of life.
Neurofilament Light Protein as a Biomarker in Severe Mental Disorders: A Systematic Review
Authors: Squitti Rosanna , Fiorenza Antonio , Martinelli Alessandra , Brembati Viviana , Crescenti Daniela , Rongioletti Mauro , Ghidoni Roberta
Published: December 2024

Severe mental disorders (SMDs), such as schizophrenia (SZ), bipolar disorder (BD), and major depressive disorder (MDD), are heterogeneous psychiatric diseases that impose a significant societal burden due to their chronic disabling nature. There are no objective and reliable diagnostic tests for SMDs; thus, there is an urgent need for specific biomarkers to improve diagnosis, treatment, and resource allocation. Neurofilaments, found in cerebrospinal fluid and blood, offer reliable diagnostic and prognostic potential. This review discusses the link between neurofilament light chain (NfL) involvement in psychiatric and neurodegenerative diseases and gives insights into the diagnostic and prognostic value of NfL in SMDs. This systematic review searched PubMed, Scopus, and Web of Science databases to answer the research question “Are NfL levels higher in individuals with SMDs compared to healthy controls?” using terms related to neurofilament, SMDs, SZ, BD, and depression. Of 8577 initial papers, 115 were relevant. After exclusions and manual additions, 17 articles were included. Studies indicate elevated NfL levels in SMDs compared to healthy controls, suggesting its potential as a biomarker for SMDs and for distinguishing neurodegenerative diseases from psychiatric disorders. However, further longitudinal research is needed to confirm its reliability for differential diagnosis, disease prediction, and treatment assessment in psychiatry.
Oxometallate-Based Ionic Liquid Catalyzed CO2-Promoted Hydration of Propargylic Alcohols for α-Hydroxy Ketones Synthesis
Authors: Wang Yuankun , Wang Chongli , Lin Weidong , Wang Qin , Li Baisong , Chen Cheng , Yuan Ye , Verpoort Francis
Published: December 2024

α-Hydroxy ketones are a crucial class of organic compounds prevalent in natural products and pharmaceutical molecules. The CO2-promoted hydration of propargylic alcohols is an efficient method for the synthesis of α-hydroxy ketones. Herein, an ionic liquid (IL) was designed to catalyze this reaction individually under atmospheric CO2 pressure, volatile organic solvents, and additives. This IL, constructed from the molybdate anion, can be recycled from industrial (NH4)2MoO4 production wastewater, demonstrating its high tolerance to catalytic environments and significant potential for practical applications. To our knowledge, this is the first instance of an oxometallate-based IL catalyst being utilized for the CO2-promoted hydration of propargylic alcohols. Further mechanistic studies revealed the bifunctionality of this IL in activating both CO2 and substrates.
Influence of Mikania micrantha Kunth Flavonoids on Composition of Soil Microbial Community
Authors: Yang Qilin , Cui Wenyang , Guan Zijun , Wang Zhenzhen , Jahan Israt , Li Ping , Qin Feng , Qiao Xi , Liu Bo , Yan Jian
Published: December 2024

Mikania micrantha, one of the world’s most destructive invasive species, is known for causing significant ecological and economic harm. While extensive research has focused on its growth characteristics, secondary metabolites, and control measures, its chemical interactions with the environment—particularly the role of flavonoids in shaping soil microbial communities—remain underexplored. In this study, we identified and quantified ten flavonoids from M. micrantha root exudates using UPLC-MS, including Hispidulin, Isorhamnetin, and Mikanin. To examine their impact, crude flavonoid extracts were applied to soil in potted experiments, which demonstrated that these compounds significantly increased soil fungal diversity and boosted the relative abundance of arbuscular mycorrhizal fungi (AMF). Furthermore, KEGG pathway analysis revealed that flavonoid addition elevated the copy numbers of genes involved in nitrogen cycling and metabolic functions, enhancing nutrient availability and microbial activity. Additionally, crude flavonoid extracts promoted the relative abundance of beneficial soil bacteria, such as Achromobacter, as well as AMF, both of which contribute to nutrient acquisition, plant growth, and soil health. These findings indicate that M. micrantha’s flavonoids can alter soil microbial community composition, thereby creating a favorable environment that reinforces its competitive edge over native plants.
The Identification and Characterization of the PeGRF Gene Family in Populus euphratica Oliv. Heteromorphic Leaves Provide a Theoretical Basis for the Functional Study of PeGRF9
Authors: Wang Ying , Wu Zhihua , Jia Mingyu , Li Jing , Song Tongrui , Jin Hongyan , Sun Jianhao , Qiu Chen , Lu Xiaona , Yuan Yang , Chen Yongqiang , Jiao Peipei , Li Zhijun
Published: December 2024

Populus euphratica Oliv. typically has four kinds of heteromorphic leaves: linear (Li), lanceolate (La), ovate (Ov) and broad ovate (Bo). Heteromorphic leaves help P. euphratica adapt to extreme desert environments and further contribute to protection against land desertification in Northwest China. In the authors’ previous research, growth-regulating factors (GRFs) were speculated to be related to the development of P. euphratica heteromorphic leaves via multi-omics integrated analysis. However, the genomic features and biological role of the P. euphratica GRF gene family in heteromorphic leaves are still unclear. In this study, 19 PeGRF genes were genome-widely identified and characterized in P. euphratica, and their physicochemical properties, gene structure and phylogenetic evolution were analyzed. An analysis of the research showed that PeGRFs were unevenly distributed on 11 chromosomes and that PeGRF proteins contained conserved motif 1 (WRC) and motif 2 (QLQ). Moreover, 19, 15, 19 and 22 GRFs were identified in Populus deltoides Marshall, Populus pruinosa Schrenk, Salix sinopurpurea C. Wang et C. Y. Yang and Salix suchowensis W. C. Cheng, respectively. A collinearity analysis showed that the PeGRF family evolved slowly within Populus species. A phylogenetic tree of the GRF family was also constructed, and GRFs were divided into four subfamilies. A large number of cis-acting elements were related to plant growth and development, plant hormone response and stress response on the promoter of PeGRFs. The expression pattern of PeGRFs showed significant up-regulation in broad leaves (Ov and Bo) compared with narrow leaves (Li and La). In combination with the predicted gene regulatory network, PeGRF9 (PeuTF06G01147.1) may have an important contribution to the leaf shape development of P. euphratica. The heterologous expression of PeGRF9 in wild-type plants (Col-0) of Arabidopsis thaliana (L.) Heynh was also studied, showing a significant increase in the leaf area of overexpressing plants compared with the wild type. Nineteen PeGRF gene members were identified and characterized in P. euphratica, and a comparison of the genomic analysis of Populus GRF members revealed their evolutionary features. The further overexpression of PeGRF9 in A. thaliana revealed its biological role in the heteromorphic leaves of P. euphratica. This study not only provides new insights into the evolution and function of PeGRFs in P. euphratica heteromorphic leaves but also helps in an understanding of the adaptive evolution of P. euphratica in drought desert environments.
Unusual Genomic and Biochemical Features of Paenarthrobacter lasiusi sp. nov—A Novel Bacterial Species Isolated from Lasius niger Anthill Soil
Authors: Dymova Alexandra A. , Kovalev Maxim A. , Silantyev Artemiy S. , Borzykh Anna A. , Osipova Pamila J. , Poddubko Svetlana V. , Mitkevich Vladimir A. , Karpov Dmitry S. , Kostina Natalia V.
Published: December 2024

The black garden ant (Lasius niger) is a widely distributed species across Europe, North America, and North Africa, playing a pivotal role in ecological processes within its diverse habitats. However, the microbiome associated with L. niger remains poorly investigated. In the present study, we isolated a novel species, Paenarthrobacter lasiusi, from the soil of the L. niger anthill. The genome of P. lasiusi S21 was sequenced, annotated, and searched for groups of genes of physiological, medical, and biotechnological importance. Subsequently, a series of microbiological, physiological, and biochemical experiments were conducted to characterize P. lasiusi S21 with respect to its sugar metabolism, antibiotic resistance profile, lipidome, and capacity for atmospheric nitrogen fixation, among others. A notable feature of the P. lasiusi S21 genome is the presence of two prophages, which may have horizontally transferred host genes involved in stress responses. P. lasiusi S21 synthesizes a number of lipids, including mono- and digalactosyldiacylglycerol, as well as steroid compounds that are typically found in eukaryotic organisms rather than prokaryotes. P. lasiusi S21 exhibits resistance to penicillins, lincosamides, fusidins, and oxazolidinones, despite the absence of specific genes conferring resistance to these antibiotics. Genomic data and physiological tests indicate that P. lasiusi S21 is nonpathogenic to humans. The genome of P. lasiusi S21 contains multiple operons involved in heavy metal metabolism and organic compound inactivation. Consequently, P. lasiusi represents a novel species with an intriguing evolutionary history, manifesting in distinctive genomic, metabolomic, and physiological characteristics. This species may have potential applications in the bioaugmentation of contaminated soils.
Synthesis of New Promising BNCT Agents Based on Conjugates of closo-Dodecaborate Anion and Aliphatic Diamino Acids
Authors: Ryabchikova Margarita N. , Nelyubin Alexey V. , Klyukin Ilya N. , Selivanov Nikita A. , Bykov Alexander Yu. , Kubasov Alexey S. , Skribitsky Vsevolod A. , Finogenova Yulia A. , Shpakova Kristina E. , Kasianov Anton A. , Lipengolts Alexey A. , Zhdanov Andrey P. , Grigoreva Elena Yu. , Zhizhin Konstantin Yu. , Kuznetsov Nikolay T.
Published: December 2024

In this work, a series of boronated amidines based on the closo-dodecaborate anion and amino acids containing an amino group in the side chain of the general formula [B12H11NHC(NH(CH2)nCH(NH3)COOH)CH3], where n = 2, 3, 4, were synthesized. These derivatives contain conserved α-amino and α-carboxyl groups recognized by the binding centers of the large neutral amino acid transporter (LAT) system, which serves as a target for the clinically applied BNCT agent para-boronophenylalanine (BPA). The paper describes several approaches to synthesizing the target compounds, their acute toxicity studies, and tumor uptake studies in vivo in two tumor models. The promising compound [B12H11NHC(NH(CH2)2CH(NH3)COOH)CH3]*3H2O demonstrates low toxicity (LD50 in a range from 150 to 300 mg/kg) and excellent solubility and also shows selective uptake in experimental melanoma in laboratory mice (T/N ratio remained >3 up to 60 min post-injection, with a maximum T/N of 6.2 ± 2.8 at 45 min).
Dual-Double Stem Cell Ovarian Therapy: A Comprehensive Approach in Regenerative Medicine
Authors: Ljubić Aleksandar , Dinić Marija , Švraka Dajana , Vujović Svetlana
Published: December 2024

Dual-double stem cell therapy, which integrates mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs), represents a cutting-edge approach in regenerative medicine, particularly for conditions such as ovarian decline, premature ovarian insufficiency (POI), and induced ovarian failure. This therapy leverages the unique properties of MSCs and HSCs, enhancing tissue repair, immune modulation, and overall regenerative outcomes. MSCs, known for their ability to differentiate into various cell types, provide a supportive microenvironment and secrete bioactive molecules that promote angiogenesis and reduce inflammation. HSCs, crucial for hematopoiesis and immune function, further enhance this environment by supporting hematopoietic processes and immune regulation. Clinical evidence increasingly supports the effectiveness of stem cell therapy in ovarian regeneration. Studies have demonstrated improved folliculogenesis, normalization of hormone profiles, and successful pregnancies in patients with POI. Furthermore, recent clinical trials in various medical fields underline the superior potential of dual-double therapy compared to monotherapies involving MSCs or HSCs alone, enhancing tissue repair and functional outcomes. However, despite these benefits, the therapy presents risks that require careful consideration. For autologous MSC therapy involving expanded cell populations, risks include tumorigenic potential, with evidence of sarcoma formation in certain cases of cultured MSCs. In contrast, autologous non-expanded MSC and HSC therapies may be limited by low cell yields, potentially compromising therapeutic efficacy. Additionally, non-expanded HSC therapy poses risks of insufficient cell numbers for successful engraftment and delayed immune reconstitution. These considerations underscore the importance of quality control and rigorous screening to optimize safety and efficacy. This article explores the mechanisms of action, clinical applications, and potential complications of dual-double stem cell therapy, underscoring the need for continued research and optimized protocols to enhance safety and outcomes in ovarian insufficiency and related conditions, offering new hope for affected women.
Roles of CDR2 and CDR2L in Anti-Yo Paraneoplastic Cerebellar Degeneration: A Literature Review
Authors: Martínez Lozada Pablo S. , Mancero Montalvo Rafael , Iturralde Carrillo Andrea , Montesdeoca-Lozada Maria , Rodas Jose A. , Leon-Rojas Jose E.
Published: December 2024

Paraneoplastic cerebellar degeneration (PCD) is a rapidly progressive, immune-mediated syndrome characterized by the degeneration of Purkinje cells, often associated with the presence of antibodies targeting intracellular antigens within these cells. These autoantibodies are implicated in the induction of cytotoxicity, leading to Purkinje cell death, as demonstrated in in vitro models. However, the precise roles of antibodies and T lymphocytes in mediating neuronal injury remain a subject of ongoing research, with T cells appearing to be the main effectors of cerebellar injury. Notably, at least 50% of PCD cases involve anti-Yo autoantibodies, also referred to as anti-PCA1 (Purkinje cell antigen 1) antibodies, which specifically target cerebellar degeneration-related protein 2 (CDR2) and its paralogue, CDR2-like (CDR2L). Another recognized antigen is CDR 34, a 34 kDa Purkinje cell antigen characterized by tandem repeats and a B-cell epitope; its detection in non-cerebellar tissues necessitates further in situ hybridization studies. Onconeural antigens are expressed in both Purkinje cells and tumour cells, where they localize in the cytoplasm and associate with membrane-bound and free ribosomes, playing critical roles in regulating transcription and calcium homeostasis. Recent studies suggest that the breakdown of immune tolerance is linked to genetic alterations in tumour cell antigens, leading to the formation of neoantigens that can elicit autoreactive T cells, which may underscore the function of Yo antibodies. In vitro studies indicate that anti-Yo antibodies can induce cell death independent of T lymphocytes. The disease progresses by initial lymphocytic infiltration, followed by a rapid loss of Purkinje cells without significant inflammation. However, in vivo models showcase that anti-Yo PCD is primarily T-cell mediated, with antibodies serving as biomarkers rather than direct effectors of neuronal death. This review examines the mechanisms underlying PCD, focusing on the roles of CDR2 and CDR2L in tumour development and their potential role in the degeneration of cerebellar Purkinje neurons. A comprehensive understanding of these processes is essential for advancing diagnostic, prognostic, and therapeutic strategies for PCD and associated malignancies.
Domain Mobility in the ORF2p Complex Revealed by Molecular Dynamics Simulations and Big Data Analysis
Authors: Kulakova Anna M. , Khrenova Maria G. , Zvereva Maria I. , Polyakov Igor V.
Published: December 2024

ORF2p (open reading frame 2 protein) is a multifunctional multidomain enzyme that demonstrates both reverse transcriptase and endonuclease activities and is associated with the pathophysiology of cancer. The 3D structure of the entire seven-domain ORF2p complex was revealed with the recent achievements in structural studies. The different arrangements of the CTD (carboxy-terminal domain) and tower domains were identified as the “closed-ring” and “open-ring” conformations, which differed by the hairpin position of the tower domain, but the structural diversity of these complexes has the potential to be more extensive. To study this, we performed sub-microsecond all-atom molecular dynamics simulations of the entire ORF2p complex with different starting configurations. The obtained molecular dynamic trajectories frames were assigned to several clusters following the dimension reduction to three principal components of the 1275 distances feature matrix. Five and six clusters were obtained for the “open” and “closed” ring models, respectively. While the fingers–palm–thumb core retains its rigid configuration during the MD (molecular dynamics) simulations, all other domains display the complicated dynamic behavior not observed in the experimental structures. The EN (endonuclease) and CTD domains display significant translations and rotations while their internal structures stay rigid. The CTD domain can either form strong contacts with the tower or be far apart from it for both formal “open” and “closed” ring states because the tower hairpin position is not the only determining factor of the protein complex configuration. While only the “thumb up” conformation is observed in all the trajectories, the active site can be obstructed by the movement of the CTD domain. Thus, molecular modeling and machine learning techniques provide valuable insights into the dynamical behavior of the ORF2p complex, which is hard to uncover with experimental methods, given the complexity and size of the object.
Can α-Mangostin and Photodynamic Therapy Support Ciprofloxacin in the Inactivation of Uropathogenic Escherichia coli and Staphylococcus aureus Strains?
Authors: Wojnicz Dorota , Korzekwa Kamila , Guźniczak Mateusz , Wernecki Maciej , Ulatowska-Jarża Agnieszka , Buzalewicz Igor , Tichaczek-Goska Dorota
Published: December 2024

Multidrug-resistant bacteria represent a significant challenge in the treatment of bacterial infections, often leading to therapeutic failures. This issue underlines the need to develop strategies that improve the efficacy of conventional antibiotic therapies. In this study, we aimed to assess whether a plant-derived compound, α-mangostin, and photodynamic therapy (PDT) could enhance the antibacterial activity of ciprofloxacin against uropathogenic strains of Escherichia coli and Staphylococcus aureus. Using nanopore sequencing technology, we confirmed that the clinical strains tested were classified as multidrug-resistant. Digital holotomography (DHT) was used to examine α-mangostin-induced changes in the bacterial cells’ penetration by a photosensitizer. A scanning confocal fluorescence microscope was used to visualize photosensitizer penetration into bacterial cells and validate DHT results. A synergistic effect between α-mangostin and ciprofloxacin was observed exclusively in S. aureus strains, while no enhancement of ciprofloxacin’s antibacterial activity was detected in E. coli strains when combined with α-mangostin. Notably, photodynamic therapy significantly potentiated the antibacterial effects of ciprofloxacin and its combination with α-mangostin compared to untreated controls. In addition, morphological changes were observed in bacterial cells exposed to these antimicrobials. In conclusion, our findings suggest that α-mangostin and PDT may serve as valuable adjuncts to ciprofloxacin, improving the eradication of uropathogens.
Quantitative Determination of a Series of Oxysterols by an Optimized LC-MS/MS Analysis in Different Tissue Types
Authors: Guo Zhiting , Yu Huiyan , Yang Kexin , Feng Wenjing , Liu Miao , Wang Tao , Xiao Rong
Published: December 2024

Oxysterols, as metabolites of cholesterol, play a key role in cholesterol homeostasis, autophagosome formation, and regulation of immune responses. Disorders in oxysterol metabolism are closely related to the pathogenesis of neurodegenerative diseases. To systematically investigate the profound molecular regulatory mechanisms of neurodegenerative diseases, it is necessary to quantify oxysterols and their metabolites in central and peripheral biospecimens simultaneously and accurately. However, there are a lot of unsolved problems with the existing methods, such as the hindrance of applying a single method to different biological specimens or the challenge of simultaneous quantification due to differential groups on the ends of the oxysterol side chains. Herein, according to the physicochemical properties and structure of oxysterols, an optimized liquid chromatography-tandem mass spectrometry method for the quantification of oxysterols was established by optimizing the sample preparation process, chromatographic conditions, mobile phase pH, and solvent selection. Seven oxysterols were detected by this method, including 27-hydroxycholesterol, 7α-hydroxycholesterol, 7α,27-dihydroxycholesterol, 7-dehydrocholesterol, 7α-hydroxy-3-oxo-4-cholestenoic acid, 3-hydroxy-5-cholestenoic acid, and 24(S)-hydroxycholesterol. Non-derivatization extraction with methyl tert-butyl ether was used for different biospecimens, followed by simultaneous chromatographic separation of oxysterols on a phenyl hexyl column. By repeated validation, this method exhibited satisfactory linearity, precision, recovery, sensitivity, repeatability, and stability, and it was successfully applied to the detection of oxysterols in the plasma, cerebral cortex, and liver of mouse. In summary, our optimized method enables concurrent analysis and quantification of oxysterols and their metabolites in various biospecimens, presenting a broad range of applicability.
Effective Targeting of Glutamine Synthetase with Amino Acid Analogs as a Novel Therapeutic Approach in Breast Cancer
Authors: Abdelsattar Shimaa , Al-Amodi Hiba S. , Kamel Hala F. , Al-Eidan Ahood A. , Mahfouz Marwa M. , El khashab Kareem , Elshamy Amany M. , Basiouny Mohamed S. , Khalil Mohamed A. , Elawdan Khaled A. , Elsaka Shorouk , Mohamed Salwa E. , Khalil Hany
Published: December 2024

Cancer cells undergo metabolic rewiring to support rapid proliferation and survival in challenging environments. Glutamine is a preferred resource for cancer metabolism, as it provides both carbon and nitrogen for cellular biogenesis. Recent studies suggest the potential anticancer activity of amino acid analogs. Some of these analogs disrupt cellular nucleotide synthesis, thereby inhibiting the formation of DNA and RNA in cancer cells. In the present study, we investigated the anticancer properties of Acivicin and Azaserine in the breast cancer MCF-7 cell line, comparing their effects to those on the non-tumorigenic MCF-10 epithelial cell line in vitro. Interestingly, at lower concentrations, both Acivicin and Azaserine showed potent inhibition of MCF-7 cell proliferation, as assessed by the MTT assay, without detectable toxicity to normal cells. In contrast, Sorafenib (Nexavar), a commonly used drug for solid tumors, showed harmful effects on normal cells, as indicated by increased lactate dehydrogenase (LDH) production in treated cells. Furthermore, unlike Sorafenib, treatment with Acivicin and Azaserine significantly affected apoptotic signaling in treated cells, indicating the role of both amino acid analogs in activating programmed cell death (PCD), as assessed by the Annexin-V assay, DAPI staining, and the relative expression of tumor suppressor genes PTEN and P53. ELISA analysis of MCF-7 cells revealed that both Acivicin and Azaserine treatments promoted the production of anti-inflammatory cytokines, including IL-4 and IL-10, while significantly reducing the production of tumor necrosis factor alpha (TNF-α). Mechanistically, both Acivicin and Azaserine treatment led to a significant reduction in the expression of glutamine synthetase (GS) at both the RNA and protein levels, resulting in a decrease in intracellular glutamine concentrations over time. Additionally, both treatments showed comparable effects on Raf-1 gene expression and protein phosphorylation when compared with Sorafenib, a Raf-1 inhibitor. Moreover, docking studies confirmed the strong binding affinity between Acivicin, Azaserine, and glutamine synthetase, as evidenced by their docking scores and binding interactions with the enzyme crystal. Collectively, these findings provide evidence for the anticancer activity of the two amino acid analogs Acivicin and Azaserine as antagonists of glutamine synthetase, offering novel insights into potential therapeutic strategies for breast cancer.
Identification of ALOX12B Gene Expression, Evolution, and Potential Functional Sites in Horn Development of Sheep
Authors: Lv Ran , Zhang Guoqing , Li Hao , Shi Jianxin , Meng Zhu , Lu Xiaoning , Shan Mingzhu , Yang Jie , Pan Zhangyuan
Published: December 2024

The growth and development of horns are primarily controlled by the skin. The ALOX12B gene is crucial for epidermal barrier function and may have a significant impact on horn growth. The purpose of this study was to investigate the expression of ALOX12B across different sheep breeds and tissues by utilizing RNA sequencing. Additionally, potential functional sites were identified in conjunction with whole genome sequencing. Our findings revealed that ALOX12B was highly expressed in the scurred horn group as opposed to the normal horn group (SHE). ALOX12B expression was also notably high in the skin across several species. Eight loci that may influence horn size were indicated in this study. Through the alignment of the ALOX12B protein sequence from 16 species, 15 amino acid sites were identified specifically expressed in horned animals. In conclusion, this study established a connection between ALOX12B and horn size and identified a series of functional sites that may serve as molecular markers for reducing the presence of horns in Chinese sheep breeds.
Total Antioxidant and Oxidative Status as Potential Biomarkers of Alcohol Overdose
Authors: Ptaszyńska-Sarosiek Iwona , Gołaś Edyta , Nesterowicz Miłosz , Niemcunowicz-Janica Anna , Zalewska Anna , Żendzian-Piotrowska Małgorzata , Maciejczyk Mateusz
Published: December 2024

Serious alcohol-associated hazards underscore the need to develop new biomarkers reflecting the biological changes caused by chronic alcohol use and predicting the risk of alcohol-related death. Oxidative stress is one mechanism of alcohol toxicity. The blood and urine redox status (total antioxidant capacity [TAC], total oxidative status [TOS], and oxidative stress index [OSI]) was assessed in 105 people who died a sudden death (controls), 47 people who died of alcohol overdose, and 102 people with alcohol dependency. TAC and TOS were determined utilizing the colorimetric method. Non-parametric tests were used for statistical analysis. Blood and urine TAC levels were significantly elevated in individuals both with alcohol dependency and alcohol poisoning compared with controls. TOS levels were elevated in the blood of both study groups compared with the control group, and significantly higher in patients with alcohol dependency compared with the group with alcohol poisoning. TAC in the blood highly correlated with blood alcohol content. Receiver operating characteristic (ROC) analysis showed that the blood TAC effectively discriminated between individuals with alcohol poisoning and alcohol dependency with high sensitivity and specificity. Our study confirmed impaired redox homeostasis in people with alcoholism and indicated the utility of TAC, TOS, and OSI as biomarkers of alcohol exposure.
Recombinant Follicle-Stimulating Hormone and Luteinizing Hormone Enhance Mitochondrial Function and Metabolism in Aging Female Reproductive Cells
Authors: Lin Li-Te , Li Chia-Jung , Lee Yi-Shan , Tsui Kuan-Hao
Published: December 2024

Ovarian aging significantly impacts female fertility, with mitochondrial dysfunction emerging as a key factor. This study investigated the effects of recombinant follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on mitochondrial function and metabolism in aging female reproductive cells. Human granulosa cells (HGL5) were treated with FSH/LH or not. Mitochondrial function was assessed through various assays, including mitochondrial mass, membrane potential, ROS levels, and ATP production. Mitochondrial dynamics and morphology were analyzed using MitoTracker staining. Cellular respiration was measured using a Seahorse Bioenergetics Analyzer. Metabolic reprogramming was evaluated through gene expression analysis and metabolite profiling. In vivo effects were studied using aging mouse oocytes. FSH/LH treatment significantly improved mitochondrial function in aging granulosa cells, increasing mitochondrial mass and membrane potential while reducing ROS levels. Mitochondrial dynamics showed a shift towards fusion and elongation. Cellular respiration, ATP production, and spare respiratory capacity were enhanced. FSH/LH-induced favorable alterations in cellular metabolism, favoring oxidative phosphorylation. In aging mouse oocytes, FSH/LH treatment improved in vitro maturation and mitochondrial health. In conclusion, FSH/LH supplementation ameliorates age-related mitochondrial dysfunction and improves cellular metabolism in aging female reproductive cells.
Non-Structural Protein V of Canine Distemper Virus Induces Autophagy via PI3K/AKT/mTOR Pathway to Facilitate Viral Replication
Authors: Tian Xin , Zhang Rui , Yi Shuang , Chen Yu , Jiang Ying , Zhang Xianwen , Zhang Zhidong , Li Yanmin
Published: December 2024

Canine distemper (CD) is a highly infectious disease of dogs which is caused by canine distemper virus (CDV). Previous studies have demonstrated that CDV infection can induce autophagy in cells. However, the mechanism underlying CDV-induced autophagy remains not fully understood. The CDV non-structural protein V plays a vital role in viral replication and pathogenicity in the host. In this study, we investigated the relationship between the CDV-V protein and autophagy induction and further explored its impact on the viral replication and the mechanism behind this. Our results showed that the V protein induced autophagy via inhibiting the phosphorylation of PI3K, AKT, and mTOR to promote viral replication. The activation or inhibition of PI3K phosphorylation resulted in enhancing or reducing viral replication, respectively. Further studies revealed that the V protein interacted with PI3K to induce cellular autophagy. The present study demonstrated that the CDV-V protein can induce cellular autophagy by inhibiting the PI3K/AKT signaling pathway to enhance viral replication. The results improve the understanding of the molecular mechanism of CDV infection and offer new perspectives for the development of effective treatment and prevention strategies.
Alpinetin Exhibits Antioxidant and Anti-Inflammatory Effects in C57BL/6 Mice with Alcoholic Liver Disease Induced by the Lieber–DeCarli Ethanol Liquid Diet
Authors: Radosavljevic Tatjana , Brankovic Milica , Djuretić Jasmina , Grujic-Milanovic Jelica , Kovacic Marijana , Jevtic Jovan , Stankovic Sanja , Samardzic Janko , Vucevic Danijela , Jakovljevic Vladimir
Published: December 2024

Alcohol-associated liver disease (ALD) is a common non-communicable chronic liver disease characterized by a spectrum of conditions ranging from steatosis and alcohol-associated steatohepatitis (AH) to fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). The pathogenesis of ALD involves a complex interplay of various molecular, biochemical, genetic, epigenetic, and environmental factors. While the mechanisms are well studied, therapeutic options remain limited. Alpinetin, a natural flavonoid with antioxidant and anti-inflammatory properties, has shown potential hepatoprotective effects, though its efficacy in ALD remains unexplored. This study investigated the hepatoprotective effects of alpinetin using a Lieber–DeCarli ethanol liquid diet model of ALD in C57BL/6 mice. Mice were divided into three groups: the control group, the ethanol group, and the ethanol group treated with alpinetin. Serum activity of ALT, AST, γ-GT, and ALP was measured to assess liver function, along with antioxidative and oxidative/nitrosative stress markers in liver tissue. Pro-inflammatory cytokines and endoplasmic reticulum (ER) stress parameters in liver tissue were also evaluated. Histological assessment of disease activity was performed using the SALVE grading and staging system. Treatment with alpinetin significantly reduced serum levels of ALT, AST, γ-GT, and oxidative/nitrosative stress markers while increasing antioxidative markers. The levels of pro-inflammatory cytokines and ER stress parameters were significantly decreased. Histological analysis demonstrated reduced steatosis, hepatocyte ballooning, and inflammation. These findings suggest that alpinetin holds promise as a potential therapeutic agent for managing ALD.
Starvation Metabolism Adaptations in Tick Embryonic Cells BME26
Authors: Nogueira Cintia Lopes , Arcanjo Angélica F. , Lima Maria Elisa , Moraes Bruno , da Silva Renato Martins , Gondim Katia C. , Konnai Satoru , Ramos Isabela , Santos Samara , Filardy Alessandra D’Almeida , Pinto Kamila Guimarães , Vaz Junior Itabajara da Silva , Logullo Carlos
Published: December 2024

Ticks are hematophagous ectoparasites that transmit pathogens and inflict significant economic losses on the cattle industry. Remarkably, they can survive extended periods of starvation in the absence of a host. The primary objective of this study was to investigate the metabolic adaptations that enable the tick Rhipicephalus microplus to endure starvation using the BME26 cell line as a model system. To simulate nutrient deprivation, cells were subjected to starvation conditions by replacing the L-15 culture medium with phosphate-buffered saline (PBS). Our findings show that these tick cells can endure experimental starvation for up to 48 h. The assessment of glycogen levels in starved cells shows a significant decrease, at both the 24 h and 48 h marks. Additionally, upregulation of phosphoenolpyruvate carboxykinase (PEPCK) gene expression, along with downregulation of hexokinase (HK) and pyruvate kinase (PK) gene expression, indicated that BME26 cells would prioritize the gluconeogenic pathway over the glycolytic pathway under starvation conditions. Moreover, the transcriptional levels of autophagy-related genes (ATG) were upregulated in response to starvation. Taken together, our findings suggest a potential role for autophagy in supplying substrates for the gluconeogenic pathway in nutrient-deprived tick cells. This work contributes to the understanding of metabolic regulation in R. microplus ticks and offers valuable insights for tick control strategies.
Profiling Genome-Wide Methylation Patterns in Cattle Infected with Ostertagia ostertagi
Authors: Boschiero Clarissa , Beshah Ethiopia , Zhu Xiaoping , Tuo Wenbin , Liu George E.
Published: December 2024

DNA methylation (DNAm) regulates gene expression and genomic imprinting. This study aimed to investigate the effect of gastrointestinal (GI) nematode infection on host DNAm. Helminth-free Holstein steers were either infected with Ostertagia ostertagi (the brown stomach worm) or given tap water only as a control. Animals were euthanized 30 days post-infection, and tissues were collected at necropsy. We conducted epigenome-wide profiling using a mammalian methylation array to explore the impact of infection on methylation patterns in the mucosa from abomasal fundus (FUN), pylorus (PYL), draining lymph nodes (dLNs), and the duodenum (DUO). The analysis covered 31,107 cattle CpGs of 5082 genes and revealed infection-driven, tissue-specific, differential methylation patterns. A total of 389 shared and 2770 tissue-specific, differentially methylated positions (DMPs) were identified in dLN and FUN, particularly in genes associated with immune responses. The shared DMPs were found in 263 genes, many of which are involved in immune responses. Furthermore, 282, 244, 52, and 24 differentially methylated regions (DMRs) were observed in dLN, FUN, PYL, and DUO, respectively. More hypomethylated DMRs were detected in dLN and FUN, while more hypermethylated DMRs were found in PYL and DUO. Genes carrying DMPs and DMRs and enriched pathways relating to immune functions/responses were detected in infected animals, indicating a link between DNA methylation and the infection. The data may implicate a crucial role of DNAm in regulating the nature/strength of host immunity to infection and contribute to a deeper understanding of the epigenetic regulatory landscape in cattle infected by GI nematodes.
The Potential of Metabolomics as a Tool for Identifying Biomarkers Associated with Obesity and Its Complications: A Scoping Review
Authors: Skowronek Anna Katarzyna , Jaskulak Marta , Zorena Katarzyna
Published: December 2024

Obesity and its related diseases, such as type 2 diabetes (T2DM), cardiovascular disease (CVD), and metabolic fatty liver disease (MAFLD), require new diagnostic markers for earlier detection and intervention. The aim of this study is to demonstrate the potential of metabolomics as a tool for identifying biomarkers associated with obesity and its comorbidities in every age group. The presented systematic review makes an important contribution to the understanding of the potential of metabolomics in identifying biomarkers of obesity and its complications, especially considering the influence of branched-chain amino acids (BCAAs), amino acids (AAs) and adipokines on the development of T2DM, MAFLD, and CVD. The unique element of this study is the combination of research results from the last decade in different age groups and a wide demographic range. The review was based on the PubMed and Science Direct databases, and the inclusion criterion was English-language original studies conducted in humans between 2014 and 2024 and focusing on the influence of BCAAs, AAs or adipokines on the above-mentioned obesity complications. Based on the PRISMA protocol, a total of 21 papers were qualified for the review and then assigned to a specific disease entity. The collected data reveal that elevated levels of BCAAs and some AAs strongly correlate with insulin resistance, leading to T2DM, MAFLD, and CVD and often preceding conventional clinical markers. Valine and tyrosine emerge as potential markers of MAFLD progression, while BCAAs are primarily associated with insulin resistance in various demographic groups. Adipokines, although less studied, offer hope for elucidating the metabolic consequences of obesity. The review showed that in the case of CVDs, there is still a lack of studies in children and adolescents, who are increasingly affected by these diseases. Moreover, despite the knowledge that adipokines play an important role in the pathogenesis of obesity, there are no precise findings regarding the correlation between individual adipokines and T2DM, MAFLD, or CVD. In order to be able to introduce metabolites into the basic diagnostics of obesity-related diseases, it is necessary to develop panels of biochemical tests that will combine them with classical markers of selected diseases.
Optimization of a Pressurized Extraction Process Based on a Ternary Solvent System for the Recovery of Neuroprotective Compounds from Eucalyptus marginata Leaves
Authors: Hasni Soumaya , Riguene Hajer , Mendiola Jose A. , Ibáñez Elena , Montero Lidia , Domínguez-Rodríguez Gloria , Ghazghazi Hanene , Rigane Ghayth , Salem Ridha Ben
Published: December 2024

Green chemistry focuses on reducing the environmental impacts of chemicals through sustainable practices. Traditional methods for extracting bioactive compounds from Eucalyptus marginata leaves, such as hydro-distillation and organic solvent extraction, have limitations, including long extraction times, high energy consumption, and potential toxic solvent residues. This study explored the use of supercritical fluid extraction (SFE), pressurized liquid extraction (PLE), and gas-expanded liquid (GXL) processes to improve efficiency and selectivity. These techniques were combined in a single mixture design, where CO2 was used in the experiments carried out under SFE, while water and ethanol were used for the PLE and GXL experiments by varying the concentration of the solvents to cover all the extraction possibilities. The neuroprotective activity of the extracts was evaluated by measuring their antioxidant, anti-inflammatory, and acetylcholinesterase inhibition properties. The optimization resulted in a novel GXL extraction with an optimal ternary mixture of 27% CO2, 55% ethanol, and 18% water, with a high degree of desirability (R2 = 88.59%). Chromatographic analysis carried out by GC-MS and HPLC-ESI-MS/MS identified over 49 metabolites. The designed sustainable extraction process offers a promising approach for producing phenolic-rich plant extracts in industrial applications.
Differential Expression of CXCL12 in Human and Mouse Hair: Androgens Induce CXCL12 in Human Dermal Papilla and Dermal Sheath Cup
Authors: Zheng Mei , An Seungchan , Park In Guk , Kim Jino , Kim Won-Serk , Noh Minsoo , Sung Jong-Hyuk
Published: December 2024

We previously demonstrated that C-X-C Motif Chemokine Ligand 12 (CXCL12) is primarily secreted by dermal fibroblasts in response to androgens and induces hair miniaturization in the mouse androgenic alopecia (AGA) model. However, the direct effects of androgen-induced CXCL12 on dermal papilla cells (DPCs) and dermal sheath cup cells (DSCs) have not been demonstrated. First, we compared single-cell RNA sequencing data between mouse and human skin, and the results show that CXCL12 is highly co-expressed with the androgen receptor (AR) in the DPCs and DSCs of only human hair. Immunohistochemistry also showed that CXCL12 is co-expressed with the AR in the DPCs and DSCs of human hair follicles. In human hair organ culture, androgens also increased CXCL12 expression in DPCs and DSCs and reduced hair length, while the CXCL12 antibody increased hair length via AR inactivation. CXCL12 mRNA was upregulated by androgen treatment in primary human DPCs and DSCs. On the contrary, AR inhibitors or siRNA treatment reduced CXCL12 expression. Collectively, these results suggest that CXCL12 is co-expressed with the AR in the DPCs and DSCs of human hair follicles; therefore, inhibition of CXCL12 using antibodies is a promising strategy for AGA treatment.
In Vitro Effect of Estrogen and Progesterone on Cytogenetic Profile of Uterine Leiomyomas
Authors: Koltsova Alla S. , Pendina Anna A. , Malysheva Olga V. , Trusova Ekaterina D. , Staroverov Dmitrii A. , Yarmolinskaya Maria I. , Polenov Nikolai I. , Glotov Andrey S. , Kogan Igor Yu. , Efimova Olga A.
Published: December 2024

In the present study, we aimed to investigate intratumoral karyotype diversity as well as the estrogen/progesterone effect on the cytogenetic profile of uterine leiomyomas (ULs). A total of 15 UL samples obtained from 15 patients were cultured in the media supplemented with estrogen and/or progesterone and without adding hormones. Conventional cytogenetic analysis of culture samples revealed clonal chromosomal abnormalities in 11 out of 15 ULs. Cytogenetic findings were presented by simple and complex chromosomal rearrangements (64% and 36% of cases, respectively) verified through FISH and aCGH. In most ULs with complex chromosomal rearrangements, the breakpoints did not feature clusterization on a single chromosome but were evenly distributed across rearranged chromosomes. The number of breakpoints showed a strong positive correlation with the number of rearranged chromosomes. Moreover, both abovementioned parameters were in a linear dependency from the number of karyotypically different clones per UL. This suggests that complex chromosomal rearrangements in ULs predominantly originate through sequential events rather than one hit. The results of UL cytogenetic analysis depended on the presence of estrogen and/or progesterone in the culture medium. The greatest variety of cytogenetically different cell clones was detected in the samples cultured without hormone supplementation. Their counterparts cultured with progesterone supplementation showed a sharp decrease in clone number, whereas such a decrease induced by estrogen or estrogen–progesterone supplementation was insignificant. These findings suggest that estrogen–progesterone balance is crucial for forming a UL cytogenetic profile, which, in turn, may underlie the unique response of the every karyotypically abnormal UL to medications.