Background/aim: Failures in the treatment of colorectal cancer (CRC) have increased scientific interest in the potential of the bioactive compounds contained in cell-free supernatants (CFSs) obtained from probiotic bacteria for cancer treatment. The aim in this study was to investigate the effects of CFSs obtained from Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) and Lactobacillus paracasei subsp. paracasei (L. paracasei) on colorectal cancer cell lines (HT-29 and Caco-2) and a healthy colon epithelial cell line (CCD841 CoN). Materials and methods: Cytotoxic effects on the cell lines were determined by MTT assay. In line with the IC50 values obtained, morphological changes were examined by fluorescent cell staining and cell cycle analyses were performed. In addition, the expression levels of the apoptosis-related genes Bax, Cas 9, Cas 8, Cas 3, and Bcl-2 in CFS-treated cells were measured using the qRT-PCR method. Results: The MTT assay revealed that CFSs exerted a significant cytotoxic effect on cancer cells. Assessments with AO/PI fluorescent staining confirmed the increase in the number of apoptotic cells. Cell cycle analyses showed a tendency for accumulation in the G0/G1 phase and a decrease in the S phase. Furthermore, in cells treated with inactive CFS, proapoptotic genes (Bax, Cas 9, Cas 8, and Cas 3) were significantly increased, while antiapoptotic Bcl-2 gene expression decreased. Conclusion: This study demonstrates that both live and inactivated Lactobacillus CFSs can exert targeted anticancer effects in Caco-2 and HT-29 cells. These results indicate that CFSs may be a potential biotherapeutic option for cancer treatment in the future.
Background/aim:Lissachatina fulica mucin (LFM) has been used as a nutraceutical, therapeutic agent, and cosmetic ingredient. The bioactivity of mucin depends on bioactive compounds whose composition is strongly influenced by the snail's diet. This study investigated the bioactive compounds in L. fulica mucin as potential therapeutic agents for atopic dermatitis (AD). Materials and methods:LFM was collected from snails fed long beans and melon rinds for 4 weeks, freeze-dried, and analyzed using liquid chromatography-mass spectrometry followed by network pharmacology (NP) analysis to identify its bioactive compounds. A total of 60 BALB/c mice were sensitized on the dorsal skin with 1% 2,4-dinitrochlorobenzene and subsequently treated with dexamethasone or LFM cream (5% or 10%), which was applied daily for 7 days. Skin samples were collected on days 0, 3, 5, and 7 for immunohistological analysis. Results:The results demonstrated that LFM contained 61 bioactive compounds, 16 of which interacted with seven target proteins: CCL2, TNF, PTGS2, SYK, MMP9, AKT1, and ICAM1. The NP analysis identified three major bioactive compounds in LFM-macamide, N-benzyloleamide, and a compound tentatively annotated as eplerenone-that potentially targeted CCL2 and TNF, which may serve as key regulators associated with the anti-AD effects of LFM. LFM 5% cream significantly reduced Atopic Dermatitis Severity Index (ADSI) scores and interleukin-1β (IL-1β) expression on day 5 (p < 0.05). LFM normalized epidermal thickness and increased the numbers of macrophages and mast cells (p < 0.05), suggesting a potential role in tissue remodeling and inflammation resolution during skin recovery. Conclusion:LFM demonstrated potential anti-AD effects by modulating CCL2- and TNF-associated pathways. LFM 5% cream reduced ADSI scores and IL-1β expression while normalizing epidermal thickness, potentially through immunomodulatory responses during the skin recovery phase.
Background/aim: Targeted therapies with monoclonal antibodies provide cancer patients with better prognosis and diseasefree survival. The blockade of immune checkpoints, including programmed cell death protein-1 (PD-1) and its ligand PD-L1, with monoclonal antibodies may boost immune responses against tumors and is regarded as an effective strategy in cancer immunotherapy. We describe the generation of anti-PD-L1 monoclonal antibodies with high affinity and specificity, and we assess their potential for therapeutic use in cancer. Materials and methods: Hybridomas were selected for PD-L1 specificity and cross-reactivity with other immune checkpoint proteins and PD-L1 orthologs using indirect ELISA. Immunofluorescence and Western blotting assays were conducted for further characterization of the antibodies. The affinities of the antibodies for PD-L1 were determined using surface plasmon resonance. Receptor blocking activities were examined through competitive ELISA and cell-based luciferase reporter assays. Sequences of variable regions of the selected antibodies were determined by Sanger sequencing and subjected to BLAST analysis. Results: A total of 25 PD-L1-specific monoclonal antibodies were generated. While most clones reacted with PD-L1 from cynomolgus monkeys, none of the antibodies displayed cross-reactivity with other checkpoint proteins. Immunofluorescence assays showed that the selected clones stained PD-L1-expressing cell membranes specifically, but not those of PD-L1-negative cells. Western blotting revealed that most of the clones recognized both glycosylated and nonglycosylated PD-L1, and a few reacted with the glycosylated form only. Only two clones with subnanomolar affinity for human PD-L1 were effective at blocking PD-1/PD-L1 and CD80/PD-L1 interactions. Sequence analysis of their variable regions revealed their unique specificity. Conclusion: Of the 25 monoclonal antibodies produced in this study, only one was identified as a potential therapeutic drug candidate thanks to its high capacity for checkpoint blockade and affinity, as well as its unique sequence specificity. These properties are comparable to those of anti-PD-L1 antibodies currently used in clinical practice.
Background/aim: WD40 repeats are found in many ciliary proteins. Although the WD40 repeat-containing WD repeat-containing protein 31 (WDR-31) is known to regulate ciliary protein trafficking and morphology, the specific contributions of its N-terminal, WD40, and C-terminal domains to protein localization and ciliary gate integrity remain unclear. Aim of this study is to dissect the functional roles of WDR-31 domains by investigating their contributions to ciliary localization, ciliary protein trafficking, and gate integrity. Materials and methods: Using CRISPR/Cas9 technology, we generated Caenorhabditis elegans strains with in-frame deletions that removed the N-terminus (WDR-31(ΔN)), WD40 (WDR-31(ΔWD)), and C-terminus (WDR-31(ΔC)). We next examined the protein dynamics of wild-type WDR-31 and these variants at the ciliary base using confocal imaging and fluorescence recovery after photobleaching. Gate function was evaluated using the distribution of the periciliary marker TRAM-1, and structural ciliary abnormalities in AWB neurons were scored using confocal microscopy. Results: When the N-terminal domain was deleted, WDR-31 became less mobile at the basal body and mislocalized to the transition zone, suggesting a role in protein turnover. Furthermore, the WD40 domain is required for basal body confinement, whereas the C-terminus prevents WDR-31 from spreading into the distal axoneme. Any single domain deletion resulted in TRAM-1 leakage into the cilium in a sensitized mutant background. Remarkably, WDR-31 localization depends on IFT, the BBSome, and transition zone components; its mobility is significantly reduced in intraflagellar transport-defective mutants, supporting a model in which its steadystate distribution at the ciliary base appears to depend on intact intraflagellar transport machinery and may involve dynamic regulation of protein localization, retention, or turnover rather than being passively restricted. Conclusion: Our results indicate that the N-terminal, WD40, and C-terminal domains of WDR-31 are necessary for its dynamic positioning at the ciliary base. Together, these domains in WDR-31 are critical for maintaining TRAM-1 exclusion and periciliary membrane compartment/ciliary membrane compartmentalization.
Background/aim:Alzheimer's disease (AD) is a neurodegenerative disorder linked to cognitive decline and memory loss, marked by hyperphosphorylated tau tangles and decreased acetylcholine levels, which are affected by acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity. Glycogen synthase kinase-3 beta (GSK-3β) also plays a role in AD through abnormal activation. In this study, the potential of β-arbutin to inhibit cholinesterases and GSK-3β was investigated, utilizing in vitro assays, cytotoxicity analysis, and molecular docking simulations to explore its therapeutic implications. Materials and methods:Ellman's technique was used to test the inhibitory effects of arbutin on cholinesterase enzymes. An ATP-Glo luminescent assay kit was used to evaluate the inhibitory effects and inhibition kinetics of arbutin on GSK-3β. A Cell Titer-Glo 2.0 assay kit was used to assess the cytotoxicity of arbutin in SH-SY5Y cells. Results:β-Arbutin exhibited significant inhibitory effects on AChE, BuChE, and GSK-3β, with IC50 values of 0.6, 4, and 1.23 μM, respectively. The inhibition kinetics of GSK-3β with respect to ATP was competitive but with respect to the substrate (GS-2) was noncompetitive. We further found that β-arbutin, when applied alone to SH-SY5Y cells, does not affect cell viability and can ensure the sustainability of cell viability and health. Docking scores were -7.980, -6.726, and -6.115 kcal/mol for GSK-3β, AChE, and BuChE, respectively. Conclusion:Our research demonstrates that arbutin is a modulator of cholinesterase and GSK-3β and may be beneficial as a multitarget-directed therapeutic natural compound for AD.
Background/aim:Cervical cancer (CC) is a malignant gynecologic tumor. Small RNAs derived from tRNAs (tsRNAs) have been reported to play regulatory roles in tumor progression and suppression. However, the functional role of tsRNAs in CC remains largely unclear. Materials and methods:Small RNA sequencing was performed on normal, cervical intraepithelial neoplasia (CIN), and CC clinical samples to identify differentially expressed tsRNAs (DEtsRNAs). qRT-PCR was used to validate the selected DEtsRNAs. The effects of tsRNA-05020 on HeLa cell proliferation, apoptosis, and epithelial-to-mesenchymal transition were assessed. Results:Compared with the normal group, 215 tsRNAs were significantly differentially expressed in CIN tissues, of which 196 were upregulated and 19 were downregulated. A total of 184 DEtsRNAs were identified between CC and CIN tissues, including 81 upregulated and 103 downregulated in CC. In total, three candidate DEtsRNAs were validated, among which tsRNA-05020 exhibited the largest fold change and was significantly downregulated in CC compared with CIN. Target gene network analysis identified 278 putative target mRNAs of tsRNA-05020 in CC. Overexpression of tsRNA-05020 significantly inhibited HeLa cell proliferation and promoted apoptosis. Moreover, overexpression of tsRNA-05020 significantly reduced vimentin expression and increased ZO-1 expression. Conclusion:This study identifies a previously uncharacterized role of tsRNA-05020 in CC progression, expands the current understanding of tsRNA-associated regulatory networks, and suggests that tsRNA-05020 may serve as a potential molecular regulator in CC.
Background/aim:Polyhydroxyalkanoates (PHAs) are biodegradable biopolymers with promising applications in biotechnology and medicine; however, the diversity of their biosynthesis within the genus Priestia remains poorly characterized. Although Priestia species have been recognized as potential PHA producers, the detailed genomic architecture and physicochemical properties of the polymers synthesized by these bacteria, particularly in the type strain, remain poorly understood. This study aimed to characterize the poly-β-hydroxybutyrate (PHB) produced by Priestia endophytica UCM B-5715 and to investigate the PHB biosynthetic genes across related strains. Materials and methods:PHB granules were visualized and measured using fluorescence microscopy, transmission electron microscopy, and scanning electron microscopy. Polymer structure and molecular weight were examined using Fourier transform infrared spectroscopy, 1H nuclear magnetic resonance spectroscopy, and viscometry, whereas thermal behavior was evaluated using differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). Comparative genomic analysis of 11 P. endophytica strains was performed to reconstruct the PHB biosynthetic pathways and to analyze the evolutionary relationships among PhaC synthases. Results:Microscopy confirmed the intracellular accumulation of PHB granules ranging in size from 65.35 × 47.77 nm to 1544.39 × 1126.53 nm. DSC analysis revealed a glass transition temperature of 2.3 °C, a crystallization temperature of 74.6 °C, and two melting peaks at 155 °C and 164.5 °C, indicating the presence of distinct crystalline domains. TGA demonstrated high thermal stability, with thermal decomposition initiating at 271 °C. Genomic analysis revealed a complete and conserved pha gene cluster across all examined strains, with the PhaC proteins being classified as Class IV PHA synthases. Conclusion:This study provides the first comprehensive physicochemical and genomic characterization of PHB synthesized by the type strain Priestia endophytica UCM B-5715, highlighting the potential of this species as a robust microbial source of biotechnologically relevant biopolymers.
Background/aim:Polyunsaturated fatty acids (PUFAs) and the hepatic cholinergic axis are critical regulators of lipid metabolism and inflammatory signaling; however, their time-dependent interplay in hepatocytes remains incompletely understood. Clarifying how n-6 and n-3 PUFAs interact with cholinergic pathways may provide mechanistic insight into metabolic dysfunction-associated steatotic liver disease (MASLD). Materials and methods:HepG2 cells were used to examine the combined effects of butyrylcholinesterase (BChE) overexpression and PUFA exposure. Cells were treated with linoleic acid (LA; n-6) or α-linolenic acid (α-LA; n-3) for 24 or 48 h. Gene expression analyses were performed to assess pathways involved in de novo lipogenesis, β-oxidation, cholinergic signaling, and inflammatory responses. Cholinesterase activities were measured enzymatically, secreted cytokines were profiled, and molecular docking simulations were performed to evaluate potential α-LA interactions with the α7 nicotinic acetylcholine receptor (α7 nAChR) and M3 muscarinic acetylcholine receptor (M3 mAChR). Results:An early antilipogenic profile and changes suggestive of increased fatty-acid oxidation (↓SREBP-1c, ↓ACC1, and ↑CPT1A) were induced by α-LA at 24 h and were partially attenuated by 48 h. LA exhibited a biphasic pattern characterized by modest early suppression followed by a pronounced proinflammatory profile at 48 h (↑TNF, ↑IL6, and ↑COX-2). BChE overexpression was associated with increased expression of lipogenic and triglyceride-related genes, whereas cotreatment with PUFAs resulted in time- and class-specific modulation. Cholinergic markers exhibited divergent expression patterns: BCHE + α-LA increased CHAT and CHRNA7 expression at 48 h, whereas ACHE mRNA expression increased under both BCHE + LA and BCHE + α-LA conditions. Enzymatic analyses demonstrated increased total cholinesterase activity in the BChE and α-LA groups, while molecular docking supported the potential accommodation of α-LA within the binding pockets of α7 nAChR and M3 mAChR. Conclusion:PUFA class and exposure duration jointly shape hepatocellular metabolic and inflammatory states, with cholinergic signaling acting as a modulatory node relevant to MASLD.
Background/aim:Inflammatory bowel disease is a chronic inflammatory disorder characterized by excessive immune activation, primarily mediated by nitric oxide and proinflammatory cytokines. This condition encompasses ulcerative colitis and Crohn's disease, both of which significantly impair patients' quality of life and pose persistent challenges in clinical management. Pectin, a complex carbohydrate found in plant cell walls, has shown promise in reducing inflammation. Materials and methods:In this study, pectin was extracted from red dragon fruit peel using two methods: ultrasound-assisted extraction and ultrasound-assisted enzymatic extraction. The structural characteristics and antiinflammatory effects of the extracted pectin were evaluated in lipopolysaccharide-stimulated RAW 264.7 macrophages. Results:Ultrasound-assisted enzymatic extraction produced pectin with a higher yield (19.38%), a lower degree of esterification (46.51%), a reduced molecular weight (94.67 kDa), and a greater galacturonic acid content (67.51%) compared with ultrasound extraction alone. Both pectin preparations were noncytotoxic to RAW 264.7 macrophages and significantly reduced nitric oxide production and the expression of key inflammatory mediators. Structural analysis confirmed the integrity of the pectin molecules. Conclusion:These findings suggest that ultrasound-assisted enzymatic extraction enhances the physicochemical properties and antiinflammatory activity of pectin, thereby supporting its potential application in future studies targeting inflammatory bowel disease.
Background/aim:Sciatic nerve injury causes a loss of skeletal muscle innervation, reduced motor function, and eventual muscle atrophy. Inflammation and increased protein degradation are key factors contributing to muscle atrophy. Inflammation is activated early during muscle atrophy and can be modulated by various factors. However, the precise role of inflammation in denervation-induced muscle atrophy remains unclear. Materials and methods:Transcriptome sequencing was used to determine that the inflammatory response occurs early during denervation-induced muscle atrophy. RT-qPCR validation of several inflammatory factors showed rapid upregulation at early stages, followed by gradual downregulation. Weighted gene coexpression network analysis of differentially expressed genes identified gene modules whose expression patterns were correlated with or inversely correlated to the inflammatory phenotype, thereby identifying key regulatory factors. A total of 14 coexpression modules were identified, and expression patterns opposite to those of inflammatory factors were examined to investigate potential regulatory molecules that could inhibit inflammation and protect skeletal muscle. Results:Ankrd2 was identified in the darkorange module, showing no significant change at 36 h postdenervation, followed by gradual upregulation, which was opposite to the expression of inflammatory factors. An Ankrd2-overexpressing lentivirus was injected into the tibialis anterior muscle, and Ankrd2 overexpression was found to significantly alleviate muscle atrophy. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that Ankrd2 overexpression was associated with downregulation of inflammation-related pathways, particularly the NF-κB signaling pathway. Proatrophy genes in both the ubiquitin-proteasome and autophagic-lysosomal systems were also suppressed. Conclusion:The present study suggests that denervation-induced muscle atrophy is alleviated by Ankrd2, potentially through inhibition of inflammation, highlighting its potential as a therapeutic target.
Background/aim:Spinal muscular atrophy (SMA) is a neurodegenerative disease caused by the loss of survival of motor neuron (SMN) protein. SMN deficiency leads to perturbations of the cytoskeleton, including microtubules, which are mainly involved in motility-related cellular processes. However, the molecular mechanisms of microtubule dysregulation in SMA remain elusive. Alpha (α)-tubulin is a structural component of microtubules, and its posttranslational modifications affect microtubule dynamics. Here, we aimed to investigate α-tubulin acetylation and related molecular mechanisms in SMA. Materials and methods:Two different SMA mouse models, the Drosophila melanogaster model and patient-derived fibroblasts, were used in the study. Western blot and quantitative microscopic analysis were performed to analyze α-tubulin acetylation and related mechanisms. Results:The acetylation level of α-tubulin was decreased in the Drosophila model and in SMA patient fibroblast cells but not in mouse models. This decrease in acetylation is associated with upregulation of the major tubulin deacetylase, HDAC6, in patient cells compared with healthy controls. Microtubules play a role in the organization of the Golgi apparatus, and we demonstrated that increasing α-tubulin acetylation by pharmacological inhibition of HDAC6 partially restored the fragmented morphology of the Golgi apparatus in SMA. Conclusion:Our findings provide new insight into the molecular basis of SMA, indicating that cellular pathologies, including abnormal Golgi morphology, are associated with microtubule dysregulations caused by altered α-tubulin posttranslational modifications and regulatory proteins. Our findings support that microtubule perturbations are part of SMA pathology.
Background:Glioblastoma (GBM) is a highly aggressive form of brain tumor characterized by rapid proliferation and invasiveness. It is associated with a poor prognosis due to acquired resistance to temozolomide (TMZ). In this study, we investigated whether a combination of epirubicin, 5-fluorouracil (5-FU), and TMZ could improve TMZ sensitivity in resistant GBM cells and help overcome resistance. Materials and methods:TMZ resistance was established in the U87MG cell line. The MTT assay was used to measure cell viability. Reactive oxygen species (ROS) and apoptosis were measured using flow cytometry. RNA-seq was used to evaluate genomic changes based on treatment with drugs alone or in combination. Results:We demonstrated that the triple-drug combination significantly reduced cell viability. The biochemical pathways involved revealed that this combination therapy significantly increased the generation of ROS. The RNA-seq analysis indicated that combination therapy effectively suppressed cell cycle regulatory pathways, enhancing cell cycle arrest and promoting apoptosis in TMZ-resistant cells. Conclusion:These findings underscore the potential viability of integrating epirubicin and 5-FU with TMZ to improve therapeutic outcomes in patients suffering from chemoresistant GBM. These combination therapies could represent an important advance in the treatment of this challenging malignancy.
Background/aim:Zearalenone (ZEA) is a Fusarium-derived mycotoxin that frequently contaminates food and feed and induces hepatotoxicity in humans and animals through oxidative stress and inflammation. Pinocembrin (PCM), a naturally occurring flavonoid, has potent antioxidant and antiinflammatory properties. This study investigated the hepatoprotective effects of PCM against ZEA-induced liver injury in albino mice, with a focus on the nuclear factor erythroid 2-related factor 2 (NRF2) and phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathways. Materials and methods:Twenty-four albino male mice were allocated into four groups: (1) control (vehicle 0.05% dimethyl sulfoxide), (2) ZEA-treated (40 mg/kg body weight), (3) PCM-treated (50 mg/kg body weight), and (4) ZEA + PCM. After 42 days of oral treatment, serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels were determined. Hepatic oxidative stress markers were assessed. Liver histopathology was evaluated using standard staining techniques. Western blot analysis was performed to determine the expression of several proinflammatory cytokines signaling proteins: interleukin-6, interleukin-1β, tumor necrosis factor-α, NRF2 pathway components, apoptosis-related proteins (B-cell lymphoma 2 [Bcl-2], cleaved caspase-3), and PI3K/Akt. Results:ZEA administration caused significantly elevated serum liver enzyme and hepatic malondialdehyde levels while causing a reduction in other oxidative stress markers. Histopathological examination revealed marked hepatic architectural disruption and cellular degeneration in the ZEA-treated group. Additionally, ZEA caused downregulation of antiapoptotic signaling proteins (such as Bcl-2) and upregulation of proinflammatory cytokines and cleaved caspase-3 expression. The coadministration of PCM facilitated markedly attenuated ZEA-induced biochemical, oxidative, inflammatory, apoptotic, and histopathological alterations. PCM treatment restored antioxidant enzyme activities and produced significant enhancement of NRF2/HO-1/NQO1 and PI3K/Akt signaling. Conclusion:PCM effectively protects against ZEA-induced hepatotoxicity in albino mice by mitigating oxidative stress, inflammation, and apoptosis. These protective effects are partially mediated through activation of the NRF2 signaling pathway and modulation of the PI3K/Akt pathway.
Background:Meiotic transcription is a highly regulated process involving stage-specific chromatin remodeling and dynamic RNA polymerase II (RNAPII) activity. Nevertheless, our current understanding is largely based on classical model organisms, while many nonmodel mammals remain unexplored. This study investigates the transcriptional dynamics and chromatin features during meiotic prophase I in Nannospalax xanthodon, a subterranean blind mole rat species with extensive intraspecific karyotype variation. Materials and methods:Using immunofluorescence, we analyzed synapsis and transcriptional activity in spermatocyte nuclei of two geographically distant N. xanthodon males, detecting SYCP3, CREST, γH2AFX, and phosphorylated RNAPII (Ser5, Ser2). Telomere-associated proteins RAP1 and TERT were also examined to assess transcriptional activity at chromosomal ends. Results:Transcriptional activity was detected at all substages of prophase I. Unlike classical models, N. xanthodon exhibited no sharp pachytene-specific transcriptional reactivation; instead, RNAPII-Ser2 and Ser5 signals increased gradually from leptotene to pachytene. Notably, RNAPII-Ser5, but not Ser2, was consistently localized at telomeric regions of autosomes and sex chromosomes. These RNAPII-Ser5 foci coincided with RAP1-positive telomeres, suggesting a poised but transcriptionally inactive polymerase state at chromosome ends. The variability of RNAPII signals around the XY bivalent may also suggest a dynamic regulation of sex chromosome silencing. This is the first analysis of meiotic transcription in mole rats. The gradual transcriptional enhancement in the chromatin of prophase I nuclei and the RNAPII-Ser5 enrichment at telomeres reflect species-specific regulatory features. Conclusions:These findings highlight the importance of investigating nonmodel organisms to uncover novel mechanisms of meiotic regulation and suggest that transcriptional control during meiosis may be more evolutionarily diverse than previously recognized.
Background/aim:Plant genetic engineering is a valuable approach for improving stress tolerance, crop productivity, and quality traits. However, its progress is limited by the drawbacks of traditional transformation methods. Commonly used techniques, such as Agrobacterium-mediated transformation and biolistics, are restricted by genotype dependency, lengthy tissue culture steps, and low efficiency, particularly in monocot species. Therefore, the development of alternative gene delivery strategies remains an ongoing research focus in plant biotechnology. Nanoparticle-based delivery systems have recently gained attention due to their physiochemical properties, biocompatibility, and potential for alternative gene delivery approaches in plant systems. Materials and methods:We investigated the use of single-walled carbon nanotubes (SWCNTs) as nanocarriers for plasmid DNA delivery in the model monocot plant Brachypodium distachyon. SWCNTs were functionalized with polyethyleneimine (PEI) and plasmid DNA (pCAMBIA1301) was loaded via electrografting. Successful complex formation was confirmed with transmission electron microscopy (TEM), atomic force microscopy (AFM), and zeta potential measurements. Results:The TEM and AFM analyses revealed that the SWCNTs retained a thin, elongated needle-like structure, consistent with their ability to pass through the plant cell wall. Zeta potential measurements showed a negative surface charge of -43 mV for COOH-SWCNTs that shifted to +56 mV after PEI functionalization, enabling electrostatic binding of plasmid DNA. pDNA-SWCNT complexes at different mass ratios were applied to both callus and seed explants to evaluate their delivery potential. Successful delivery was confirmed by b-glucuronidase (GUS) reporter gene expression in both callus tissues and seeds, with ImageJ-based quantification showing that the 3:1 SWCNT:pDNA ratio yielded the highest mean signal intensity. Conclusion:Our findings show that the SWCNT-based system enabled the delivery of a large plasmid (~12 kb), supporting transient reporter gene expression in monocot tissues. Overall, the data suggest that SWCNT-mediated delivery represents a promising technique for DNA delivery in monocots, and further optimization is expected to improve its efficiency.
This corrigendum is to address an issue regarding the manuscript’s previous publication. The authors noticed that the representative flow cytometry image for the AAA group in Figure 3B, as well as the representative Histone H3 and β-actin Western blot bands in Figure 4A, were incorrectly used in the original published version of this paper and indicated that the relevant corrections do not affect the results or the conclusions of the study.To rectify this oversight and ensure the accuracy of the published work, the correct figures are included for your reference. Link to the original article: https://doi.org/10.55730/1300-0152.2754
The Editors of Turkish Journal of Biology wish to issue an Expression of Concern regarding the article entitled: “Characterization of TFIIE-regulated genes by transcriptome analysis” by Serdar Baysal published on 20.12.2024. Following the publication of this article, concerns have been raised regarding• Potential violations of academic ethical standards in the published study• Potential intellectual property infringement, including plagiarism• An academic sanction reportedly imposed on the author by the affiliated university following an institutional disciplinary investigation. An investigation is currently underway to assess the validity of these concerns. In accordance with the guidelines of the Committee on Publication Ethics (COPE), and to maintain transparency with our readership, we are issuing this Expression of Concern while the matter is being reviewed. This notice does not imply conclusive evidence of misconduct or error at this stage but serves to alert readers to potential issues affecting the reliability of the article. Further editorial action will be taken as appropriate once the investigation is complete. We will update this notice in due course based on the outcome of the investigation. Link to the original article: https://doi.org/10.55730/1300-0152.2718
Background/aim: The present study investigates the role of Shenfu injection in the treatment of yang-deficient chronic heart failure (CHF). Materials and methods: Sprague-Dawley (SD) rats were modeled for yang-deficient CHF by abdominal aortic coarctation. Echocardiography was performed to detect changes in cardiac function, and serum N-terminal B-type natriuretic peptide proteins (NT-proBNP), cardiac troponin I (cTnI), and ferroptosis-related factors were measured using ELISA kits. Pathological changes in cardiac tissues were observed through hematoxylin-eosin (HE) and Masson' trichrome staining, cardiomyocyte apoptosis was measured by TUNEL staining, and reactive oxygen species (ROS) production was determined through dihydroethidium (DHE) staining. The expression of nuclear factor E2-related factor 2 (Nrf2), cyclooxygenase 2 (Ptgs2), glutathione peroxidase 4 (GPX4), solute carrier family 3 member 2 (SLC3A2), solute carrier family 7 member 11 (SLC7A11), and acyl-CoA synthetase long-chain family member 4 (ACSL4) in cardiac tissues were analyzed through RT-qPCR. Phosphorylated Akt (p-Akt), phosphorylated GSK-3(3 (p-GSK-3(3), and Nrf2 expression in tissues were tested through immunohistochemistry. The protein expression of the Akt/GSK-3(3/Nrf2 pathway was detected by Western blot. The Akt/GSK-3(3/Nrf2 pathway inhibitor LY294002 was applied to the rats administrated with Shenfu injection. Results: Shenfu injection decreased the left ventricular end-diastolic diameter and left ventricular end-systole diameter and increased the left ventricular ejection fraction and left ventricular fractional shortening in rats with CHF. The treatment reduced NT-proBNP and cTnI levels, while improving pathological damage in the cardiac tissue. The treatment was also noted to decrease serum MDA, ACSL4, and Fe2+ and increase GSH, GPX4, SOD, and SLC3A2 in the sample; increase GPX4,SLC7A11 and SLC3A2 mRNA in cardiac tissues, and decrease Ptgs2 and ACSL4 mRNA. Shenfu injection was also noted to activate the Akt/GSK-3(3/Nrf2 signaling pathway, while LY294002 weakened the therapeutic effect of the treatment on cardiac tissue damage. Conclusion: Shenfu injection activates the Akt/GSK-3(3/Nrf2 pathway to prevent myocardial injury and ferroptosis in yang-deficient CHF.
Background/aim:Breast cancer remains a major malignancy among women, and severe side effects and the development of acquired drug resistance frequently hinder current therapeutic strategies. The Notch signaling pathway, a key regulator of cell fate, is commonly dysregulated in breast cancer and associated with poor prognosis. Gamma-secretase inhibitors (GSIs) block Notch receptor activation and have shown potential anticancer efficacy. This study aimed to investigate the synergistic activity of two commonly used GSIs, DAPT and MK0752, combined with docetaxel or cisplatin in both 2D and 3D breast cancer models. Materials and methods:Triple-negative, highly metastatic MDA-MB-231 and ER+/PR+ MCF-7 breast cancer cell lines were treated with DAPT or MK0752 alone or in combination with docetaxel or cisplatin. Drug efficacy and potential synergism were evaluated in 2D monolayer cultures and 3D spheroid models. Sequential treatment strategies were also assessed, where docetaxel or cisplatin was administered prior to GSI exposure. Results:Both MDA-MB-231 and MCF-7 cell lines exhibited notable sensitivity to DAPT and MK0752 combinations with docetaxel or cisplatin in 2D and 3D cultures. Synergistic enhancement of cytotoxicity was observed, particularly in sequential treatment regimens. Pretreatment with docetaxel or cisplatin followed by GSI exposure demonstrated superior growth inhibition compared with either monotherapy or simultaneous combination treatments. Conclusion:This study highlights the therapeutic potential of combining GSIs with standard chemotherapeutics to overcome drug resistance in breast cancer. The observed synergy and sequencing effects provide a strong basis for further mechanistic and translational investigations to optimize GSI-based combinational therapy strategies.
Background/aim:Glioblastoma multiforme (GBM) is one of the most aggressive and fatal malignancies of the central nervous system. Despite advancements in treatment strategies, effective therapies for GBM remain insufficient, necessitating further improvements. Notably, miR-22 has been found to be significantly downregulated in both glioblastoma tissues and cell lines. In this study, we aim to evaluate miR-22 expression levels in GBM (U87) and CD133-positive (CD133+) GBM stem cells (GSCs) and to investigate its effects on proliferation, colony formation, migration, invasion, and wound-healing in U87 and CD133+ U87 cells in vitro. Materials and methods:We isolated CD133+ U87 cells using magnetic-activated cell sorting and determined the percentage of CD133+ cells by flow cytometry. qRT-PCR detected miR-22 expression. We transfected miR-22 miRNA into U87, CD133+, and CD133- U87 cells using a lipid-based transfection reagent. Cell viability was assessed spectrophotometrically on days 1, 3, 5, and 7 using the CCK-8 viability assay. Transwell assays were used to analyze migration and invasion. Wound healing was assessed using a scratch assay. Results:MiR-22 expression was lower in CD133+ U87 cells than in U87 cells. MiR-22 overexpression suppressed proliferation in U87, CD133+, and CD133- U87 cells. MiR-22 overexpression also inhibited migration and invasion in both CD133+ and CD133- U87 cells and impaired wound-healing capacity in both U87 and CD133- U87 cells. Conclusion:These results suggest that miR-22 acts as a tumor suppressor in GBM and CD133+ GSCs. Therefore, miR-22 represents a potential therapeutic target for cancer stem cell-based glioblastoma treatment.
