bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Analysis of the Chloroplast Genomes of the Melliodendron (Styracaceae) Species: Providing Insights into Molecular Evolution and Phylogenetic Relationships
Authors: Dai Wei , Zheng Haozhi , Xu Menghan , Zhu Xingli , Long Hui , Xu Xiaogang , Fang Yanming
Published: December 2024

Melliodendron xylocarpum is a member of the Styracaceae family, which is well-known for its remarkable ornamental and medicinal properties. In this research, we conducted comparative analysis of the chloroplast genomes from four samples of M. xylocarpum, representing Melliodendron. The results demonstrated that the chloroplast genome of four M. xylocarpum samples ranging from 157,103 bp to 158,357 bp exhibited a typical quadripartite structure, including one large single-copy (LSC) region (90,131 bp to 90,342 bp), one small single-copy (SSC) region (18,467 bp to 18,785 bp), and two inverted repeat regions (IRs) (24,115 bp to 24,261 bp). Different levels of expansion and contraction were observed in the IR region of four M. xylocarpum samples. Besides, accD and ycf1 have been identified under positive selection, potentially linked to the adaptive response of Melliodendron to various environmental changes. Conflicting phylogenetic relationships were identified among various genera within the Styracaceae family in the phylogenetic tree constructed using CDS sequences and complete chloroplast genomes. Furthermore, the significance of a large sample size was also highlighted in this study for enhancing the accuracy of findings from phylogenetic analyses. The findings of this research will provide significant insights for future investigations into the evolutionary trends and conservation of the Melliodendron species.
Characterization and Biomedical Applications of Electrospun PHBV Scaffolds Derived from Organic Residues
Authors: Jin Anyi , Pérez Germán , Martínez de Ilarduya Antxon , del Valle Luis J. , Puiggalí Jordi
Published: December 2024

This study explores the characterization and application of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) synthesized from organic residues, specifically milk and molasses. Six PHBV samples with varying 3-hydroxyvalerate (3HV) content (7%, 15%, and 32%) were analyzed to assess how 3HV composition influences their properties. Comprehensive characterization techniques, including NMR, FTIR, XRD, DSC, TGA, and tensile-stress test, were used to evaluate the molecular structure, thermal properties, crystalline structure, and mechanical behavior. Selected PHBV samples were fabricated into nanofibrous scaffolds via electrospinning, with uniform fibers successfully produced after parameter optimization. The electrospun scaffolds were further analyzed using DSC, GPC, and SEM. Biological evaluations, including cytotoxicity, in vitro drug release, and antibacterial activity tests, were also conducted. The results indicate that the electrospun PHBV scaffolds are biocompatible and exhibit promising properties for biomedical applications such as tissue engineering and drug delivery. This study demonstrates the potential of using organic residues to produce high-value biopolymers with tailored properties for specific applications.
Comparative Genomic Analysis Reveals Key Changes in the Genome of Acremonium chrysogenum That Occurred During Classical Strain Improvement for Production of Antibiotic Cephalosporin C
Authors: Zhgun Alexander A.
Published: December 2024

From the 1950s to the present, the main tool for obtaining fungal industrial producers of secondary metabolites remains the so-called classical strain improvement (CSI) methods associated with multi-round random mutagenesis and screening for the level of target products. As a result of the application of such techniques, the yield of target secondary metabolites in high-yielding (HY) strains was increased hundreds of times compared to the wild-type (WT) parental strains. However, the events that occur at the molecular level during CSI programs are still unknown. In this paper, an attempt was made to identify characteristic changes at the genome level that occurred during CSI of the Acremonium chrysogenum WT strain (ATCC 11550) and led to the creation of the A. chrysogenum HY strain (RNCM F-4081D), which produces 200–300 times more cephalosporin C, the starting substance for obtaining cephalosporin antibiotics of the 1st–5th generations. We identified 3730 mutational changes, 56 of which led to significant disturbances in protein synthesis and concern: (i) enzymes of primary and secondary metabolism; (ii) transporters, including MDR; (iii) regulators, including cell cycle and chromatin remodeling; (iv) other processes. There was also a focus on mutations occurring in the biosynthetic gene clusters (BGCs) of the HY strain; polyketide synthases were found to be hot spots for mutagenesis. The obtained data open up the possibility not only for understanding the molecular basis for the increase in cephalosporin C production in A. chrysogenum HY, but also show the universal events that occur when improving mold strains for the production of secondary metabolites by classical methods.
Molecular Sentinels: Unveiling the Role of Sirtuins in Prostate Cancer Progression
Authors: Chouhan Surbhi , Muhammad Naoshad , Usmani Darksha , Khan Tabish H. , Kumar Anil
Published: December 2024

Prostate cancer (PCa) remains a critical global health challenge, with high mortality rates and significant heterogeneity, particularly in advanced stages. While early-stage PCa is often manageable with conventional treatments, metastatic PCa is notoriously resistant, highlighting an urgent need for precise biomarkers and innovative therapeutic strategies. This review focuses on the dualistic roles of sirtuins, a family of NAD+-dependent histone deacetylases, dissecting their unique contributions to tumor suppression or progression in PCa depending on the cellular context. It reveals their multifaceted impact on hallmark cancer processes, including sustaining proliferative signaling, evading growth suppressors, activating invasion and metastasis, resisting cell death, inducing angiogenesis, and enabling replicative immortality. SIRT1, for example, fosters chemoresistance and castration-resistant prostate cancer through metabolic reprogramming, immune modulation, androgen receptor signaling, and enhanced DNA repair. SIRT3 and SIRT4 suppress oncogenic pathways by regulating cancer metabolism, while SIRT2 and SIRT6 influence tumor aggressiveness and androgen receptor sensitivity, with SIRT6 promoting metastatic potential. Notably, SIRT5 oscillates between oncogenic and tumor-suppressive roles by regulating key metabolic enzymes; whereas, SIRT7 drives PCa proliferation and metabolic stress adaptation through its chromatin and nucleolar regulatory functions. Furthermore, we provide a comprehensive summary of the roles of individual sirtuins, highlighting their potential as biomarkers in PCa and exploring their therapeutic implications. By examining each of these specific mechanisms through which sirtuins impact PCa, this review underscores the potential of sirtuin modulation to address gaps in managing advanced PCa. Understanding sirtuins’ regulatory effects could redefine therapeutic approaches, promoting precision strategies that enhance treatment efficacy and improve outcomes for patients with aggressive disease.
The Role of GFAP in Post-Mortem Analysis of Traumatic Brain Injury: A Systematic Review
Authors: Sacco Matteo Antonio , Gualtieri Saverio , Tarallo Alessandro Pasquale , Verrina Maria Cristina , Calafiore Jasmine , Princi Aurora , Lombardo Stefano , Ranno Francesco , Di Cello Alessandro , Gratteri Santo , Aquila Isabella
Published: December 2024

Traumatic brain injuries (TBIs) are a leading cause of mortality and morbidity, particularly in forensic settings where determining the cause of death and timing of injury is critical. Glial fibrillary acidic protein (GFAP), a biomarker specific to astrocytes, has emerged as a valuable tool in post-mortem analyses of TBI. A PRISMA-based literature search included studies examining GFAP in human post-mortem samples such as brain tissue, cerebrospinal fluid (CSF), serum, and urine. The results highlight that GFAP levels correlate with the severity of brain injury, survival interval, and pathological processes such as astrocyte damage and blood–brain barrier disruption. Immunohistochemistry, ELISA, and molecular techniques were commonly employed for GFAP analysis, with notable variability in protocols and thresholds among studies. GFAP demonstrated high diagnostic accuracy in distinguishing TBI-related deaths from other causes, particularly when analyzed in CSF and serum. Furthermore, emerging evidence supports its role in complementing other biomarkers, such as S100B and NFL, to improve diagnostic precision. However, the review also identifies significant methodological heterogeneity and gaps in standardization, which limit the generalizability of findings. Future research should focus on establishing standardized protocols, exploring biomarker combinations, and utilizing advanced molecular tools to enhance the forensic application of GFAP.
Expression of Lumican and Osteopontin in Perivascular Areas of the Glioblastoma Peritumoral Niche and Its Value for Prognosis
Authors: Salinas María Dolores , Rodriguez Pablo , Rubio Gonzalo , Valdor Rut
Published: December 2024

Glioblastoma (GB) is one of the most aggressive and treatment-resistant cancers due to its complex tumor microenvironment (TME). We previously showed that GB progression is dependent on the aberrant induction of chaperone-mediated autophagy (CMA) in pericytes (PCs), which promotes TME immunosuppression through the PC secretome. The secretion of extracellular matrix (ECM) proteins with anti-tumor (Lumican) and pro-tumoral (Osteopontin, OPN) properties was shown to be dependent on the regulation of GB-induced CMA in PCs. As biomarkers are rarely studied in TME, in this work, we aimed to validate Lumican and OPN as prognostic markers in the perivascular areas of the peritumoral niche of a cohort of GB patients. Previously, we had validated their expression in GB xenografted mice presenting GB infiltration (OPN) or GB elimination (Lumican) dependent on competent or deficient CMA PCs, respectively. Then, patient sample classification by GB infiltration into the peritumoral brain parenchyma was related to GB-induced CMA in microvasculature PCs, analyzing the expression of the lysosomal receptor, LAMP-2A. Our results revealed a correlation between GB-induced CMA activity in peritumoral PCs and GB patients’ outcomes, identifying three degrees of severity. The perivascular expression of both immune activation markers, Iba1 and CD68, was related to CMA-dependent PC immune function and determined as useful for efficient GB prognosis. Lumican expression was identified in perivascular areas of patients with less severe outcome and partially co-localizing with PCs presenting low CMA activity, while OPN was primarily found in perivascular areas of patients with poor outcome and partially co-localizing with PCs presenting high CMA activity. Importantly, we found sex differences in the incidence of middle-aged patients, being significantly higher in men but with worse prognosis in women. Our results confirmed that Lumican and OPN in perivascular areas of the GB peritumoral niche are effective predictive biomarkers for evaluating prognosis and monitoring possible therapeutic immune responses dependent on PCs in tumor progression.
Harnessing Mesenchymal Stromal Cells for Advanced Wound Healing: A Comprehensive Review of Mechanisms and Applications
Authors: Nasadiuk Khrystyna , Kolanowski Tomasz , Kowalewski Cezary , Wozniak Katarzyna , Oldak Tomasz , Rozwadowska Natalia
Published: December 2024

Chronic wounds and injuries remain a substantial healthcare challenge, with significant burdens on patient quality of life and healthcare resources. Mesenchymal stromal cells (MSCs) present an innovative approach to enhance tissue repair and regeneration in the context of wound healing. The intrinsic presence of MSCs in skin tissue, combined with their roles in wound repair, ease of isolation, broad secretory profile, and low immunogenicity, makes them especially promising for treating chronic wounds. This review explores the current landscape of MSC application, focusing on preclinical and clinical data across chronic wounds, diabetic ulcers, burns, non-union bone fractures, lower extremity venous ulcers, pressure ulcers, and genetic skin conditions like epidermolysis bullosa. Special emphasis is given to the mechanisms through which MSCs exert their regenerative effects, underscoring their potential in advancing wound healing therapies and supporting the broader field of regenerative medicine.
Detection of Human GPCR Activity in Drosophila S2 Cells Using the Tango System
Authors: Salim Emil , Hori Aki , Matsubara Kohei , Takano-Shimizu Toshiyuki , Pratomo Andre Rizky , Marianne Marianne , Syahputra Armia , Husori Dadang Irfan , Inoue Asuka , Abdullah Maryam Aisyah , Shamsudin Nur Farisya , Rullah Kamal , Kuraishi Takayuki
Published: December 2024

G protein-coupled receptors (GPCRs) are essential cell surface proteins involved in transducing extracellular signals into intracellular responses, regulating various physiological processes. This study validated the use of the Tango assay, a sensitive method for detecting GPCR activation, in Drosophila Schneider 2 (S2) cells, focusing on the human Dopamine Receptor D4 (DRD4). Plasmids encoding the LexA-tagged human DRD4 receptor and a luciferase reporter were co-transfected into Drosophila S2 cells and stimulated with dopamine. Receptor activation was measured by quantifying the luciferase activity. The system showed high specificity for dopamine, with no activation in response to octopamine, a non-ligand for DRD4. Furthermore, the system effectively detects activation by a novel compound. These results demonstrate that Drosophila S2 cells, coupled with the Tango assay, provide a viable model for studying human GPCR function and ligand specificity. This system enables the rapid screening of potential GPCR ligands in a cost-effective cellular model.
Flecainide Specifically Targets the Monovalent Countercurrent Through the Cardiac Ryanodine Receptor, While a Dominant Opposing Ca2+/Ba2+ Current Is Present
Authors: Gaburjakova Jana , Domsicova Michaela , Poturnayova Alexandra , Gaburjakova Marta
Published: December 2024

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a highly arrhythmogenic syndrome triggered by stress, primarily linked to gain-of-function point mutations in the cardiac ryanodine receptor (RyR2). Flecainide, as an effective therapy for CPVT, is a known blocker of the surface-membrane Na+ channel, also affecting the intracellular RyR2 channel. The therapeutic relevance of the flecainide-RyR2 interaction remains controversial, as flecainide blocks only the RyR2 current flowing in the opposite direction to the physiological Ca2+ release from the sarcoplasmic reticulum (SR). However, it has been proposed that charge-compensating countercurrent from the cytosol to SR lumen plays a critical role, and its reduction may indeed suppress excessive diastolic SR Ca2+ release through RyR2 channels in CPVT. Monitoring single-channel properties, we examined whether flecainide can target intracellular pathways for charge-balancing currents carried by RyR2 and SR Cl− channels under cell-like conditions. Particularly, the Tris+ countercurrent flowed through the RyR2 channel simultaneously with a dominant reverse Ca2+/Ba2+ current. We demonstrate that flecainide blocked the RyR2-mediated countercurrent without affecting channel activity. In contrast, the SR Cl− channel was completely resistant to flecainide. Based on these findings, it is reasonable to propose that the primary intracellular target of flecainide in vivo is the RyR2-mediated countercurrent.
Atrial Fibrosis in Atrial Fibrillation: Mechanistic Insights, Diagnostic Challenges, and Emerging Therapeutic Targets
Authors: Karakasis Paschalis , Theofilis Panagiotis , Vlachakis Panayotis K. , Korantzopoulos Panagiotis , Patoulias Dimitrios , Antoniadis Antonios P. , Fragakis Nikolaos
Published: December 2024

Atrial fibrosis is a hallmark of atrial cardiomyopathy and plays a pivotal role in the pathogenesis of atrial fibrillation (AF), contributing to its onset and progression. The mechanisms underlying atrial fibrosis are multifaceted, involving stretch-induced fibroblast activation, oxidative stress, inflammation, and coagulation pathways. Variations in fibrosis types—reactive and replacement fibrosis—are influenced by patient-specific factors such as age, sex, and comorbidities, complicating therapeutic approaches. The heterogeneity of fibrosis leads to distinct electrophysiological abnormalities that promote AF via reentrant activity and enhanced automaticity mechanisms. Despite advancements in imaging, such as late gadolinium enhancement CMR and electroanatomical mapping, challenges in accurately quantifying fibrosis persist. Emerging therapeutic strategies include antifibrotic agents targeting the renin–angiotensin–aldosterone system, novel pathways like TGF-β signaling, and cardio-metabolic drugs like SGLT2 inhibitors and GLP-1 receptor agonists. Innovative interventions, including microRNA modulation and lipid nanoparticle-based therapies, show promise but require validation. Knowledge gaps remain in correlating clinical outcomes with fibrosis patterns and optimizing diagnostic tools. Future research should focus on precise phenotyping, integrating advanced imaging with molecular biomarkers, and conducting robust trials to evaluate antifibrotic therapies’ efficacy in reducing AF burden and related complications.
Differential Expression of Proteins Involved in Skin Barrier Maintenance and Vitamin D Metabolism in Atopic Dermatitis: A Cross-Sectional, Exploratory Study
Authors: Grieco Teresa , Paolino Giovanni , Moliterni Elisa , Chello Camilla , Sernicola Alvise , Egan Colin Gerard , Morelli Mariangela , Nannipieri Fabrizio , Battaglia Santina , Accoto Marina , Tirotta Erika , Trasciatti Silvia , Bonaretti Silvano , Pellacani Giovanni , Calvieri Stefano
Published: December 2024

Atopic dermatitis (AD) is a chronic inflammatory skin disorder influenced by proteins involved in skin barrier maintenance and vitamin D metabolism. Using an intra-patient design, this study compared protein expression in intra-lesional (IL) and peri-lesional (PL) skin biopsies from AD patients and examined associations between protein levels, vitamin D status, and clinical features. Forty-four biopsies from twenty-two AD patients were analyzed using antibody microarrays targeting twelve proteins. IL samples had significantly higher total protein levels than PL samples, with a mean difference of 77.7% (p < 0.001). Several proteins, including cathelicidin, cingulin, occludin, filaggrin, and the vitamin D receptor, were upregulated in IL samples. Patients with vitamin D levels below 30 ng/mL showed higher expression of CYP24A (p = 0.054), alpha-catenin (p = 0.043), and haptoglobin (p = 0.033). Increased EASI scores (≥16) were associated with elevated expression of CYP24A (p = 0.024), CYP27B (p = 0.044), filaggrin (p = 0.027), occludin (p = 0.049), and claudin-1 (p = 0.052). Multivariate regression analysis identified significant correlations between protein expression, skin prick test positivity, and low vitamin D levels. These findings suggest that proteins related to epithelial barrier function and vitamin D metabolism are highly upregulated in IL skin regions, offering potential therapeutic targets for improving both skin barrier function and overall disease severity in AD patients.
Prolonged Hypoxia in Rat Living Myocardial Slices Affects Function, Expression, and Structure
Authors: Waleczek Florian J. G. , Cipriano Giuseppe , Haas Jonas A. , Garg Ankita , Pfanne Angelika , Just Annette , Neumüller Susanne , Hegermann Jan , Pich Andreas , Radocaj Ante , Xiao Ke , Weber Natalie , Thum Thomas
Published: December 2024

Ischemic heart disease is the leading cause of death worldwide. Reduced oxygen supply and myocardial hypoxia lead to tissue damage and impairment of the heart function. To the best of our knowledge, the primary functional effects of hypoxia in the multicellular model of living myocardial slices (LMSs) have not been investigated so far. In this study, we analyzed force generation, ultrastructure, gene expression, and proteome changes in rat LMS after 24 h of ex vivo culture in normal and reduced levels of oxygen (O2). We observed a significant reduction in absolute force and a slowdown of force kinetics as well as an increase in cardiomyocyte apoptosis and myofibrillar and mitochondrial damage, as well as transcriptomic changes. Proteome analysis revealed the deregulation of proteins involved in metabolic processes, hypoxic response, and neutralizing of reactive oxygen species. Our results indicate that hypoxia induces substantial primary changes in heart tissue, which are independent of perfusion and immune responses. Our new LMS model could serve as a screening system for drug development and new mechanistic insights.
Roles of Mature Domain Targeting Signals (MTSs) for Protein Translocation and Secretion in Lactococcus lactis
Authors: Hoang Mai Ngoc , Peterbauer Clemens
Published: December 2024

Lactococcus lactis is a potential bacterial cell factory to develop delivery systems for vaccines and therapeutic proteins. Much progress has been made in applications using engineered L. lactis against, e.g., inflammatory bowel disease and cervical cancer, but the improvement of secretion and cell anchoring efficacy is still desirable. A double-labeling method based on biarsenical hairpin binding and nickel–polyhistidine affinity was used for visualization of protein trafficking and the quantification of targeted proteins on the cell surface and in the cytoplasm. To investigate the importance of mature domain targeting signals (MTSs), we generated truncated constructs encoding 126, 66, and 26 amino acid residues from the N-terminus of the basic membrane protein A (BmpA) and fused those with the gene for the human papillomavirus serotype 16 (HPV16) E7 oncoprotein. Overexpression of fusion proteins was observed to come at the cost of cell proliferation. L. lactis cells produced and displayed the shortest fusion protein only with difficulty, suggesting that the entire absence of a homologous sequence containing MTSs significantly impedes the export and surface anchoring of fusion proteins. With 40 amino acids following the signal peptide and containing one MTS, effective translocation was possible. Mutations of MTSs towards increased hydrophobicity resulted in increased secreted and surface-displayed fusion protein, suggesting the potential to design rationally improved constructs.
An Overview of the Current State of Cell Viability Assessment Methods Using OECD Classification
Authors: Madorran Eneko , Ambrož Miha , Knez Jure , Sobočan Monika
Published: December 2024

Over the past century, numerous methods for assessing cell viability have been developed, and there are many different ways to categorize these methods accordingly. We have chosen to use the Organisation for Economic Co-operation and Development (OECD) classification due to its regulatory importance. The OECD categorizes these methods into four groups: non-invasive cell structure damage, invasive cell structure damage, cell growth, and cellular metabolism. Despite the variety of cell viability methods available, they can all be categorized within these four groups, except for two novel methods based on the cell membrane potential, which we added to the list. Each method operates on different principles and has its own advantages and disadvantages, making it essential for researchers to choose the method that best fits their experimental design. This review aims to assist researchers in making this decision by describing these methods regarding their potential use and providing direct references to the cell viability assessment methods. Additionally, we use the OECD classification to facilitate potential regulatory use and to highlight the need for adding a new category to their list.
Local Application of Minimally Manipulated Autologous Stromal Vascular Fraction (SVF) Reduces Inflammation and Improves Bilio-Biliary Anastomosis Integrity
Authors: Klabukov Ilya , Shatveryan Garnik , Bagmet Nikolay , Aleshina Olga , Ivanova Elena , Savina Victoria , Gilmutdinova Ilmira , Atiakshin Dmitry , Ignatyuk Michael , Baranovskii Denis , Shegay Peter , Kaprin Andrey , Eremin Ilya , Chardarov Nikita
Published: December 2024

Bilio-biliary anastomosis (BBA) is a critical surgical procedure that is performed with the objective of restoring bile duct continuity. This procedure is often required in cases where there has been an injury to the extrahepatic bile ducts or during liver transplantation. Despite advances in surgical techniques, the healing of BBA remains a significant challenge, with complications such as stricture formation and leakage affecting patient outcomes. The stromal vascular fraction (SVF), a heterogeneous cell population derived from adipose tissue, has demonstrated promise in regenerative medicine due to its rich content of stem cells, endothelial progenitor cells, and growth factors. The objective of this study was to evaluate the potential of locally administered autologous SVF to enhance the healing of BBAs. Bilio-biliary anastomosis was performed on a swine model (female Landrace pigs). Six swine were divided into two groups: the treatment group (n = 3) received a local application of autologous SVF around the anastomosis site immediately following BBA formation, while the control group (n = 3) received saline. The primary outcomes were assessed over an eight-week period post-surgery, and included anastomosis healing, stricture formation, and bile leakage. Histological analysis was performed to evaluate fibrosis, angiogenesis, and inflammation. Immunohistochemistry was conducted to assess healing-related markers (CD34, α-SMA) and the immunological microenvironment (CD3, CD10, tryptase). The SVF-treated group exhibited significantly enhanced healing of the BBA. Histological examination revealed increased angiogenesis and reduced fibrosis in the SVF group. Immunohistochemical staining demonstrated higher vascular density in the anastomosed area of the SVF-treated group (390 vs. 210 vessels per 1 mm2, p = 0.0027), as well as a decrease in wall thickness (1.9 vs. 1.0 mm, p = 0.0014). There were no statistically significant differences in mast cell presence (p = 0.40). Immunohistochemical staining confirmed the overexpression of markers associated with tissue repair. Local injections of autologous SVF at the site of BBA have been demonstrated to significantly enhance healing and promote tissue regeneration. These findings suggest that SVF could be a valuable adjunctive therapy in BBA surgery, potentially improving surgical outcomes. However, further investigation is needed to explore the clinical applicability and long-term benefits of this novel approach in clinical practice as a minimally manipulated cell application.
Therapeutic Efficacy and Promise of Human Umbilical Cord Mesenchymal Stem Cell-Derived Extracellular Vesicles in Aging and Age-Related Disorders
Authors: Zhang Anyuan , Li Qiubai , Chen Zhichao
Published: December 2024

The global issue of aging populations has become increasingly prominent, thus the research and development for anti-aging therapies to assure longevity as well as to ameliorate age-related complications is put high on the agenda. The young humoral milieu has been substantiated to impart youthful characteristics to aged cells or organs. Extracellular vesicles (EVs) are a heterogeneous group of cell-derived membrane-limited structures that serve as couriers of proteins and genetic material to regulate intercellular communication. Of note, EVs appeared to be an indispensable component of young blood in prolonging lifespans, and circulating EVs have been indicated to mediate the beneficial effect of a young milieu on aging. Human umbilical cord mesenchymal stem cell-derived EVs (HUCMSC-EVs), isolated from the youngest adult stem cell source, are speculated to reproduce the function of circulating EVs in young blood and partially revitalize numerous organs in old animals. Robust evidence has suggested HUCMSC-EVs as muti-target therapeutic agents in combating aging and alleviating age-related degenerative disorders. Here, we provide a comprehensive overview of the anti-aging effects of HUCMSC-EVs in brain, heart, vasculature, kidney, muscle, bone, and other organs. Furthermore, we critically discuss the current investigation on engineering strategies of HUCMSC-EVs, intending to unveil their full potential in the field of anti-aging research.
The Use of a Novel Container for Secured Transport and Storage of Biological Material for Quantitative Human RNA Analysis
Authors: Kostrzewa-Nowak Dorota , Trzeciak-Ryczek Alicja , Lewandowska Klaudyna , van de Wetering Thierry , Ciechanowicz Andrzej , Nowak Robert
Published: December 2024

The transport of biological materials must protect samples from degradation and ensure courier safety. The main goal of this study was to evaluate the usefulness of a new type of container designed for the secured transport of biological material for storing samples for quantitative RNA analyses. This was achieved by analyzing changes in the expression of selected human leucocyte housekeeping genes (ACTB, GAPDH, and Rack1) using reverse transcription quantitative PCR (RT-qPCR) and digital PCR (RT-dPCR) techniques. Digital PCR analysis evidenced that the novel type of container retains a higher count of analyzed gene copies per µL of samples during 5 h of incubation time. The container ensures a low maintenance temperature for several hours, making it useful for sustaining the conditions for transporting biological samples. This novel container can be used to store and transport biological material to be analyzed by molecular techniques and can retain the stability of total RNA over several hours.
Development of Peptide Mimics of the Human Acetylcholine Receptor Main Immunogenic Region for Treating Myasthenia Gravis
Authors: Trinh Vu B. , Fairclough Robert H.
Published: December 2024

We have designed and produced 39 amino acid peptide mimics of the Torpedo and human acetylcholine receptors’ (AChRs) main immunogenic regions (MIRs). These conformationally sensitive regions consist of three non-contiguous segments of the AChR α-subunits and are the target of 50–70% of the anti-AChR autoantibodies (Abs) in human myasthenic serum and in the serum of rats with a model of that disease, experimental autoimmune myasthenia gravis (EAMG), induced by immunizing the rats with the Torpedo electric organ AChR. These MIR segments covalently joined together bind a significant fraction of the monoclonal antibodies (mAbs) raised in rats against electric organ AChR. Many of these mAbs cross react with the rat neuromuscular AChR MIR and induce myasthenic symptoms when injected into naïve rats. The human MIR mimic peptide (H39MIR) is evolutionarily related to that of the Torpedo electric organ MIR mimic peptide (T39MIR) with eight amino acid differences between the two MIR mimics. The mAbs raised to the electric organ AChR MIR cross react with the human and scores of other species’ neuromuscular AChRs. However, the mAbs do not cross react with the H39MIR mimic attached to the N-terminus of an intein–chitin-binding domain (H39MIR-IChBD) even though they do bind to the T39MIR-IChBD construct. To account for this difference in binding anti-MIR mAbs, each of the eight human amino acids was substituted individually into the T39MIR-IChBD, and four of them were found to weaken mAb recognition. Substituting the corresponding four Torpedo amino acids individually and in combination into the homologous positions in H39MIR-IChBD makes chimeric human MIR mimic peptides (T/H39MIR), some of which bind anti-MIR mAbs and anti-MIR Abs from rat EAMG and human MG sera. The best mAb binding chimeric peptide constructs may potentially serve as the basis of a diagnostic anti-MIR Ab titer assay that is both prognostic and predictive of disease severity. Furthermore, the best peptides may also serve as the targeting element of a non-steroidal antigen-specific treatment of MG to remove anti-AChR MIR Abs, either as fused to the N-terminals of the human immunoglobin Fc fragment or as the targeting component of a T cell chimeric autoantibody receptor (CAAR) directed to anti-MIR memory B cells for elimination.
Innovative Therapeutic Strategies for Myocardial Infarction Across Various Stages: Non-Coding RNA and Stem Cells
Authors: Zhuang Bingqi , Zhong Chongning , Ma Yuting , Wang Ao , Quan Hailian , Hong Lan
Published: December 2024

Myocardial infarction (MI) is a highly challenging and fatal disease, with diverse challenges arising at different stages of its progression. As such, non-coding RNAs (ncRNAs), which can broadly regulate cell fate, and stem cells with multi-differentiation potential are emerging as novel therapeutic approaches for treating MI across its various stages. NcRNAs, including microRNAs (miRNAs) and long non-coding RNAs (LncRNAs), can directly participate in regulating intracellular signaling pathways, influence cardiac angiogenesis, and promote the repair of infarcted myocardium. Currently, stem cells commonly used in medicine, such as mesenchymal stem cells (MSCs) and induced pluripotent stem cells (iPSCs), can differentiate into various human cell types without ethical concerns. When combined with ncRNAs, these stem cells can more effectively induce directed differentiation, promote angiogenesis in the infarcted heart, and replenish normal cardiac cells. Additionally, stem cell-derived exosomes, which contain various ncRNAs, can improve myocardial damage in the infarcted region through paracrine mechanisms. However, our understanding of the specific roles and mechanisms of ncRNAs, stem cells, and exosomes secreted by stem cells during different stages of MI remains limited. Therefore, this review systematically categorizes the different stages of MI, aiming to summarize the direct regulatory effects of ncRNAs on an infarcted myocardium at different points of disease progression. Moreover, it explores the specific roles and mechanisms of stem cell therapy and exosome therapy in this complex pathological evolution process. The objective of this review was to provide novel insights into therapeutic strategies for different stages of MI and open new research directions for the application of stem cells and ncRNAs in the field of MI repair.
Stem Cell-Based Therapies for Glaucoma Treatment: A Review Bridging the Gap in Veterinary Patients
Authors: de Sousa Moreira Alícia , Lopes Bruna , Sousa Ana Catarina , Coelho André , Sousa Patrícia , Araújo Ana , Delgado Esmeralda , Alvites Rui , Maurício Ana Colette
Published: December 2024

Retinal diseases are characterized by progressive damage to retinal cells, leading to irreversible vision loss. Among these, glaucoma stands out as a multifactorial neurodegenerative disease involving elevated intraocular pressure, retinal ganglion cell apoptosis, and optic nerve damage, ultimately resulting in blindness in both humans and dogs. Stem cell-based therapies have emerged as a promising therapeutic option for such conditions due to their regenerative and neuroprotective potential. These therapies, particularly those based on mesenchymal stem cells, offer the potential to repair and protect retinal tissues through the bioactive molecules (growth factors, cytokines, chemokines) secreted, their secretome. However, research in this field, especially on the use of umbilical cord mesenchymal stem cells’ secretome, remains sparse. Most clinical trials focus on human glaucomatous patients, leaving a significant gap in veterinary patients’ application, especially in dogs, with additional research being needed to determine its usefulness in canine glaucoma treatment. Future studies should aim to evaluate these therapies across both human and veterinary contexts, broadening treatment possibilities for glaucoma.
Localization of Melanocortin 1 Receptor in the Substantia Nigra
Authors: Ehara Ayuka , Ito Nozomi , Nakadate Kazuhiko , Tokuda Nobuko
Published: December 2024

Recent findings have revealed that melanocortin 1 receptor (MC1R) deficiency leads to Parkinson’s disease-like dopaminergic neurodegeneration in the substantia nigra (SN). However, its precise distribution and expressing-cell type in the SN remain unclear. Therefore, in this study, we analyzed the localization and characteristics of MC1R in the SN using histological methods, including in situ hybridization and immunohistochemistry. Our findings reveal that MC1R was slightly present in dopaminergic neurons in the ventral tier of SN pars compacta dorsal (vSNCD), a region particularly vulnerable to PD-related neurodegeneration. Notably, we discovered that MC1R is highly present in parvalbumin (PV)-positive neurons, which were also vesicular GABA transporter messenger RNA-expressing inhibitory neurons of the lateral SN pars reticulata (lSNR). Intracellular analysis demonstrated that MC1R was present not only in the plasma membrane but also in mitochondrial and endoplasmic reticulum membranes. Furthermore, MC1R co-localized with attractin (Atrn), a known MC1R modulator, in nearly all MC1R-positive neurons. Therefore, it has been suggested that MC1R and Atrn work together to regulate dopaminergic neurons in the SN through both direct expression and indirect modulation via PV-positive inhibitory neurons. These findings provide new insights into MC1R’s role in the SN and its potential contribution to PD pathophysiology
Structural Characterization and Immune Activation Capacity of Peptidoglycan from Corynebacterium glutamicum in RAW264.7 Cells
Authors: Wang Xiaoying , Li Shuzhen , Zheng Aijuan , Chen Zhimin , Chen Jiang , Zou Zhiheng , Liu Guohua
Published: December 2024

Peptidoglycan (PGN) is a unique component of prokaryotic cell walls with immune-enhancing capacities. Here, we extracted PGN from Corynebacterium glutamicum, a by-product of amino acid fermentation, using the trichloroacetic acid (TCA) method. SDS-PAGE analysis confirmed the presence of PGN, with a band of approximately 28 kDa. Further analysis was conducted through amino acid analysis, FTIR, and MALDI-TOF/TOF MS, and the results showed that the chemical structural monomer of PGN is NAG-(β-1,4-)-NAM-l-Ala-d-Glu-l-Lis-d-Ala. The immune activation effects of PGN were evaluated in a RAW264.7 cell model. Our results showed that PGN could increase the secretion level of NO, ROS, and immune regulatory substances, including TNF-α and IL-1β, and up-regulated the mRNA expression of TNF-α and iNOS. In addition, PGN stimulated the expression of ERK2, MyD88, RIP2, and the related receptor NOD1 in the NF-κB and MAPK pathways. Comparative RNA sequencing was conducted to analyze the gene expression profiles in RAW264.7 cells. KEGG analysis indicated that most of the genes were enriched in the NF-κB, MAPK, and TNF signaling pathways. Taken together, these findings suggest that PGN may have immune-activating potential for the development and application of immune adjuvants. Importantly, the application of PGN also provides a new way to utilize amino acid fermentation by-products.
Clove Essential Oil as a Source of Antitumoral Compounds Capable of Crossing the Blood–Brain Barrier: A Focus on the Effects of β-Caryophyllene and Eugenol in a Glioblastoma Cell Line
Authors: Spigarelli Renato , Spisni Enzo , Magalhães Mariana , Cabral Célia , Gonçalves Ana Cristina , Saracino Ilaria Maria , Botti Giada , Dalpiaz Alessandro , Beggiato Sarah , Valerii Maria Chiara
Published: December 2024

This study aimed to investigate β-Caryophyllene (BCA) pharmacokinetics as well as the potential antitumor activity and mechanism of action of BCA and eugenol (EU), alone or in combination, in U87 glioblastoma (GB) cells. The BCA pharmacokinetic was studied by evaluating its concentration profiles in rat blood and cerebrospinal fluid after oral and intravenous administration. EU and BCA antitumor mechanisms were assessed by comparing their effects in U87 GB cells and non-tumoral HMC3 cells. Cell death, cell cycle regulation and mitochondrial membrane potential (MMP) were evaluated using flow cytometry. mRNA levels of target genes were evaluated by qPCR. Secreted cytokines were measured by Luminex®. BCA, as well as EU, permeates the brain. EU and BCA affected the viability and proliferation of U87 cells (up to 50%, p < 0.001) but not HMC3 cells and showed a synergistic effect. BCA and EU induced G0/G1 cell cycle arrest, increasing apoptosis/necrosis. EU and BCA induced the downregulation of mRNAs encoding for key proteins involved in GB angiogenesis (VEGFA decreased op to 60%, p < 0.01), proliferation and progression, and showed anti-inflammatory activity (IL-4 significantly decreased, p < 0.001). EU and BCA demonstrated strong and multitarget antitumor activity in U87 cells. Our results provide a strong rationale for the further evaluation of EU and BCA as possible therapeutic molecules in GB management.
Nutritional Strategies for Enhancing Performance and Training Adaptation in Weightlifters
Authors: Hwang Dong-Joo , Yang Hong-Jun
Published: December 2024

Weightlifting demands explosive power and neuromuscular coordination in brief, repeated intervals. These physiological demands underscore the critical role of nutrition, not only in optimizing performance during competitions but also in supporting athletes’ rigorous training adaptations and ensuring effective recovery between sessions. As weightlifters strive to enhance their performance, well-structured nutritional strategies are indispensable. In this comprehensive review, we explored how weightlifters can optimize their performance through targeted nutritional strategies, including carbohydrate intake for glycogen replenishment and proteins for muscle growth and recovery. Additionally, the roles of key supplements, such as creatine, beta-alanine, and branch-chained amino acids in enhancing strength, delaying fatigue, and supporting muscle repair were discussed. A comprehensive literature review was conducted using PubMed, Google Scholar, and Web of Science to gather studies on nutritional strategies for weightlifting performance and training adaptation. The review focused on English-language articles relevant to weightlifters, including studies on powerlifting, while excluding those involving non-human subjects. Weightlifting requires explosive power, and proper nutrition is vital for performance and recovery, emphasizing the role of carbohydrate, protein, and fat intake. Nutrient timing and personalized strategies, informed by genetic and metabolomic analyses, enhance recovery and performance, while supplements like creatine, caffeine, and beta-alanine can significantly improve results when used correctly. Sustainable nutritional strategies are essential for enhancing weightlifter performance, emphasizing a balanced approach over extreme diets or excessive supplements. Further research is needed to refine these strategies based on individual athlete characteristics, ensuring consistent top-level performance throughout competitive seasons.
Traditional Chinese Medicine Monomer Bakuchiol Attenuates the Pathogenicity of Pseudomonas aeruginosa via Targeting PqsR
Authors: Zeng Jing , Ma Xin , Zheng Yu , Liu Dandan , Ning Wanqing , Xiao Wei , Mao Qian , Bai Zhenqing , Mao Renjun , Cheng Juanli , Lin Jinshui
Published: December 2024

As the antibiotic resistance of pathogens becomes increasingly severe, it is becoming more feasible to use methods that suppress the virulence of pathogens rather than exerting selective pressure on their growth. Pseudomonas aeruginosa, a dangerous opportunistic pathogen, infects hosts by producing multiple virulence factors, which are regulated by quorum-sensing (QS) systems, including the las systems, rhl systems, and pqs systems. This study used the chromosome lacZ transcription fusion reporter model to screen the traditional Chinese medicine monomer library and found that bakuchiol can effectively inhibit the pqs system and related virulence phenotypes of P. aeruginosa, including the production of virulence factors (pyocyanin, hydrogen cyanide, elastase, and lectin) and motility (swarming, swimming, and twitching motility) without affecting its growth. Subsequently, through genetic complementation analysis, we found that bakuchiol inhibited the function of the transcriptional activation protein PqsR of the pqs system in P. aeruginosa in a concentration-dependent manner. Furthermore, molecular dynamics simulation study results indicated that bakuchiol can target PqsR of the pqs system, thereby inhibiting the pqs system. Among the amino acids in PqsR, ALA-168 may be a key amino acid residue in the hydrophobic interaction between PqsR protein and bakuchiol. Finally, in vivo experiments demonstrated that bakuchiol attenuated the pathogenicity of P. aeruginosa to Chinese cabbage (Brassica pekinensis) and Caenorhabditis elegans. In summary, this study suggests that bakuchiol is an effective inhibitor that targets the pqs system of P. aeruginosa, providing a new strategy for addressing P. aeruginosa infections.
The Regulatory Role of Pancreatic Enzymes in the Maintenance of Small Intestinal Structure and Enterocyte Turnover with Special Reference to Alpha Amylase
Authors: Zaworski Kamil , Wychowański Piotr , Szkopek Dominika , Woliński Jarosław , Donaldson Janine , Pierzynowski Stefan , Pierzynowska Kateryna
Published: December 2024

The aim of this study was to elucidate the impact of porcine pancreatic enzymes (Creon® pancrelipase) in comparison to microbial-derived alpha amylase (MD amylase) on the small intestine wall structure, mucosal glycogen accumulation, and enterocyte turnover. The impact of enzyme supplementation on the small intestine was explored in 18 pigs with surgically induced exocrine pancreatic insufficiency (EPI). Four healthy pigs served as the control group. EPI led to reduced villus length, crypt depth, and thickness of the mucosa and muscularis layers compared to those of healthy pigs. All these changes appeared to be reversible after enzyme supplementation. Brush border thickness was decreased in EPI and increased with both enzyme preparations, with MD amylase treatment leading to the highest values in the proximal jejunum. No EPI-induced changes were observed in the goblet cell (GC) population, but significant increases in GC number and area were observed following MD amylase treatment. Glycogen accumulation within the duodenal mucosa was significantly increased in EPI pigs. EPI was also shown to significantly increase apoptotic activity and decrease proliferative activity in comparison to healthy animals, while both enzyme preparations resulted in the complete recovery of both proliferative and apoptotic activity in all investigated intestinal segments. Creon® influenced the morphology of the small intestine. However, supplementation of exogenous microbial amylase alone also affected gut morphology in a similar way to that of the complex host pancreatic enzymes offered orally. These data indicate that in addition to their role in digestion of nutrients in EPI, intraluminal pancreatic enzymes, especially amylase, contribute to gut health through maintenance of the intestinal wall architecture and physiological enterocyte turnover.