DAPI Research Articles

A new free radical scavenger, resorcimoline, prevents ischaemia/reperfusion injury in the heart

Abstract Introduction Ischaemic heart disease remains the leading cause of death worldwide. Early coronary revascularisation is the most important treatment. Recent findings have revealed that reactive oxygen species (ROS) can lead to tissue damage following coronary artery revascularisation, known as ischaemia-reperfusion injury. In this study, we focused on resorcimoline, a new free radical scavenger that was generated during the synthesis of dimethylresveratrol and dimethylpiceatannol. Last year, we reported the scavenging activity in vitro against multiple ROS, confirmed by the electron spin resonance spectrometry (Ref. 1). Purpose This study aimed to assess the potential of resorcimoline as a novel therapeutic treatment for reducing the degree of infarction caused by myocardial ischaemia, by eliminating ROS generated during myocardial ischaemia-reperfusion. Methods Nine-to-eleven-week-old male Wistar rats were subjected to acute myocardial ischaemia induced by ligation of the left anterior descending coronary artery for 30 minutes, followed by reperfusion for 24 hours. Rats received intravenous injections of resorcimoline at 6 mg/kg before coronary ligation or saline as a control (n = 9 vs. n = 8). The heart sections were double-stained with Evans blue 1% and triphenyltetrazolium chloride 1% to assess infarct area and area-at-risk. For immunohistochemical study, the other ischaemia/reperfusion models were treated with resorcimoline and saline (n = 7 vs. n = 7) as above, and the middle parts of the hearts were embedded in optimal cutting temperature compound and quickly frozen. In the H＆E-stained section, the infarct border zone was defined as a 2-mm band surrounding the necrosis area. Troponin I levels in serum were measured using ELISA. Primary cultured cardiomyocytes were treated with 10 μM angiotensin II for 3.5 hours and then exposed to resorcimoline for 30 minutes at an increasing dosage. A cellular ROS assay kit was used to assess the levels of ROS in the cardiomyocytes and the number of cells was counted with DAPI stain using an image processing software. Results Infarct size in resorcimoline-treated animals was significantly smaller than in control animals (41.8±19.4% vs. 60.8±12.9%, p=0.03, Fig. 1A) and area-at-risk was not significantly different (58.0±27.8% vs. 51.9±18.5%, p=0.61). Apoptotic cells in the border zone were significantly reduced in resorcimoline-treated animals (32.0±11.7% vs. 55.1±16.9%, p=0.01, Fig. 1B). Troponin I levels in resorcimoline-treated animals were significantly lower than in control animals (4524±3596 vs. 7856±4653, p=0.04, Fig. 1C). resorcimoline significantly reduced angiotensin II-induced ROS in cardiomyocytes in a concentration-dependent manner (Fig. 2A,B). Conclusion This novel compound, resorcimoline, can reduce infarct size induced by coronary ischaemia-reperfusion injury by reducing the ROS damage to myocardiocytes in the border zone and cellular ROS levels in cardiomyocytes.

Unraveling the Ferroptosis-inducing Potential of Methanol Leaves Extract of Prosopis Juliflora Via Downregulation of SLC7A11 and GPX4 mRNA Expression in A549 Lung Cancer Cells.

Prosopis juliflora has been employed in many traditional treatments. As evidenced by our earlier research, Prosopis juliflora leaf methanol extract (PJME) has a promising future in the fight against lung cancer. It may also be used in conjunction with other treatments to effectively manage lung cancer. The main objective of this study was to explore the potential of PJME to inhibit lung cancer in A549 cells, along with its underlying mechanisms of action. The antiproliferative effects were determined using MTT and LDH tests. Apoptosis- inducing capacity was evaluated using the DAPI staining, caspase-3 test, cytochrome C assay, PARP cleavage, and qRT-PCR. To investigate the mechanism of action of PJME in lung cancer, the levels of ROS, MMP, GSH, MDA, and specific ferroptosis indicators were measured. The experimental data of the current study indicated that exposure of A549 cells to PJME reduced cell viability and increased cellular cytotoxicity. The apoptosis-inducing ability of PJME in A549 cells was validated by enhanced nuclear condensation, level of the caspase- 3, cytochrome C, and PARP release. In addition, qRT-PCR investigations verified that the administration of PJME led to a decrease in the expression of anti-apoptotic gene Bcl2 while enhancing the mRNA level of pro-apoptotic genes, such as Bax and caspase-3, in A549 cells. The study also found that PJME has the ability to activate ferroptosis pathways, as evidenced by elevated reactive oxygen species (ROS) generation, changes in the levels of antioxidant markers (MDA and GSH), and decreased expression of SLC7A11 and GPX4. The results of the present study clearly showed that PJME inhibited the proliferation of A549 cells and induced ferroptosis by reducing the expression of the important targets SLC7A11 and GPX4. Further research is necessary to fully understand the clinical efficacy of PJME before it can be investigated as supplemental or adjuvant therapy for lung cancer.

Experimental study on repairing rat abdominal wall defect with chitosan hydrogel/polypropylene mesh composite

To investigate the improvement effects and mechanisms of composite chitosan (CS) hydrogel on traditional polypropylene (PP) mesh for repairing abdominal wall defects. CS hydrogel was prepared via physical cross-linking and then combined with PP mesh to create a CS hydrogel/PP mesh composite. The internal structure and hydrophilicity of the composite were characterized using macroscopic observation, upright metallographic microscope, scanning electron microscopy, and water contact angle measurements. The performance of the composite (experimental group) in resisting cell adhesion and supporting cell infiltration was assessed through fibroblast (NIH-3T3) infiltration experiments and human umbilical vein endothelial cells (HUVECs) tube formation assays, and simple cells were used as control group. Finally, a bilateral abdominal wall defect model (1.5 cm×1.0 cm) was established in 18 Sprague Dawley rats aged 8-10 weeks, with the composite used on one side (experimental group) and PP mesh on the other side (control group). The effects on promoting wound healing, preventing adhesion, angiogenesis, and anti-inflammation were investigated through macroscopic observation, histological staining (HE and Masson staining), and immunohistochemical staining (CD31, CD68). The composite appeared as a pale yellow, transparent solid with a thickness of 2-3 mm, with the PP mesh securely encapsulated within the hydrogel. Scanning electron microscopy revealed that the hydrogel contained interconnected pores measuring 100-300 μm, forming a porous structure. Contact angle measurements indicated that CS hydrogel exhibited good hydrophilicity, while PP mesh was highly hydrophobic. In vitro cell culture experiments showed that DAPI staining indicated fewer positive cells in the experimental group after 1 day of culture, while the cells in control group covered the entire well plate. After 3 days of culture, the cells in experimental group were spherical and displayed uneven fluorescence, suggesting that the material could reduce cell adhesion while supporting cell infiltration. HUVECs tube formation experiments demonstrated an increase in cell numbers in experimental group with a trend towards tube formation, while cells in control group were sparsely distributed and showed no migration. In the rat abdominal wall defect repair experiment, results showed that after 1 week post-surgery, the experimental group had tissue and blood vessels infiltrating, and by 4 weeks, the integrity was well restored with significant regeneration of muscle and blood vessels, while the control group exhibited adhesions and incomplete healing. HE staining results indicated weaker cell infiltration in the experimental group, with cell density significantly higher than that of the control group at 2 and 4 weeks post-surgery ( P<0.05). Masson staining revealed that collagen fibers in the experimental group were arranged neatly, with significantly increased collagen content at 2 weeks post-surgery ( P<0.05), while collagen content was similar in both groups at 4 weeks ( P>0.05). Immunohistochemical staining showed that CD31-positive cells were evenly distributed between muscle layers in the experimental group, whereas the control group exhibited notable defects. At 2 weeks after operation, the CD31-positive cell ratio was significantly higher than that in the control group ( P<0.05); at 2 and 4 weeks after operation, the CD68-positive cell ratio in the experimental group was significantly lower than that in the control group ( P<0.05). CS hydrogel has a positive effect on preventing adhesions and promoting wound healing, exhibiting anti-inflammatory and pro-angiogenic properties during the healing process. This provides a promising strategy to address challenges related to abdominal adhesions and reconstruction.

Mechanism research of Tollip negative feedback regulation in TLR4 signaling pathways based on spinal tuberculosis: Detection of Tollip and NF-κB expression levels

The emergence of drug-resistant mycobacterium tuberculosis (MTB, or TB) strains has led to an increasing incidence of TB. Spinal tuberculosis is the most common extrapulmonary tuberculosis. In the present study, tollip, a negative feedback regulatory factor in TLR4 signaling pathway was chosen based on previous studies on osteoarticular tuberculosis. U937 cells were transfected with recombinant lentivirus containing shRNA (RNA interference, RNAi) or overexpression vector containing Tollip gene and tested in vitro. The expression levels of Tollip and TLR4 were detected by Real-time PCR and immunofluorescence techniques, and the cell morphology and infection effect were observed by DAPI staining. The results suggested that Tollip gene could negatively inhibit the expression of related factors in TLR4 signaling pathway, and thus is a potential biomarker for early diagnosis.

Design, Development of Pyrazole-Linked Spirocyclopropyl Oxindole-Carboxamides as Potential Cytotoxic Agents and Type III Allosteric VEGFR-2 Inhibitors.

Tumor progression depends on angiogenesis, which is stimulated by growth factors like VEGF, targeting VEGFR kinase with small molecules is an effective anti-angiogenic therapeutic approach. The rational modification of sunitinib (VEGFR-2 inhibitor) to spirocyclopropyloxindoline carboxamides have been performed and their in vitro cytotoxic profiling was evaluated. The molecular modelling studies enabled the screening of designed analogues and identifying the possible interactions within the type III allosteric inhibitor binding site of VEGFR-2. The biological screening of synthesized compounds 15 a-y, revealed the ability of compound 15 w to inhibit the cell growth in MCF-7 cell line with IC50 value of 3.87±0.19 μM and alongside inhibition of VEGFR-2 kinase at a IC50 concentration of 4.34±0.13 μM was observed. Also, VEGFR-2 inhibition was validated through HUVEC tube formation inhibition assay. The qualitative assessment of apoptosis induction by 15 w in MCF-7 cells was evaluated through staining studies such as AO/EB and DAPI staining, whereas quantification of apoptosis and cell cycle analysis were performed through FACS analysis. The metastatic ability of the cancer cells was evaluated through inhibition of cell migration by a scratch wound healing assay. The current study strives to sequentially optimize the structural attributes of the 3-alkenyl oxindole core to surpass the existing challenges of well-known VEGFR-2 inhibitors. The findings observed from this study highlights that compound 15 w to be a prominent lead towards the development of clinical drug candidates.

PAMAM dendrimers based co-delivery of methotrexate and berberine for targeting of Hela cancer cells

Polyamidoamine dendrimer (PAMAM) is a class of synthetic macromolecular polymers for targeted drug delivery. PAMAM dendrimers are characterized by a pure defined structure, adjustable nanoscale dimensions, mono-dispersity, and versatile surface modification. The objective of this study was to study the covalent coupling of methotrexate (MTX) to PAMAM dendrimer, which was loaded with the natural product anticancer agent, berberine (BER) for the targeting of HeLa cells. The successful preparation of MTX-conjugated PAMAM loaded with BER (MTX-PAMAM-BER) was confirmed by Fourier transform infrared spectroscopy and particle size was evaluated using dynamic light scattering. The biological assays, MTT, flow cytometry, ROS levels evaluation and DAPI staining were used to assess the cytotoxicity effect of the prepared nanosystem. The findings indicated that MTX-PAMAM-BER exhibited greater suppression of tumor cell growth in comparison to BER, MTX, PAMAM-BER, and MTX-PAMAM. Besides, the noteworthy ROS level has been seen in the treated cells with MTX-PAMAM-BER. Finally, it should be stated that the fabricated MTX-PAMAM-BER co-delivery nanosystem could be a promising agent for cancer therapy and targeting.

Folate-Functionalized Chitosan-PLGA Nanoparticles: A Novel approach for targeted osthole delivery in pancreatic cancer

Efficient drug delivery systems targeting cancer cells are crucial for enhancing cancer therapy. In this study, we developed PLGA nanoparticles coated with folate-conjugated chitosan (osthole-PLGA-NPs/CS-FA) to deliver osthole to cancer cells and investigated its inhibitory and molecular signaling mechanisms in the PANC-1 pancreatic cancer cell line. Field emission scanning electron microscopy (FESEM) revealed that osthole-PLGA-NPs/CS-FA had a spherical structure with a uniform size distribution. Dynamic light scattering (DLS) analysis showed an average size of 171.76 nm, a dispersion index 0.26, and a surface charge of + 33.08 mV, indicating stability and uniform dispersion. Fourier-transform infrared (FTIR) spectrum analysis confirmed the successful incorporation of osthole into the PLGA nanoparticles, with an encapsulation efficiency of 93.12 %. These physicochemical properties suggest efficient cellular uptake and targeted delivery. The antioxidant potential of osthole-PLGA-NPs/CS-FA was evaluated using the ABTS assay, showing concentration-dependent inhibition of free radicals with an IC50 value of 172.95 μg/mL. The anticancer properties were assessed using the MTT assay, demonstrating a significant and concentration-dependent cytotoxic effect on PANC-1 cells (IC50 = 31.2 μg/mL) with minimal impact on normal human foreskin fibroblast (HFF) cells. DAPI staining and flow cytometry analyses confirmed a concentration-dependent increase in apoptosis in PANC-1 cells. The nanoparticles induced upregulation of Bax and downregulation of Bcl2, indicating activation of the intrinsic mitochondrial apoptotic pathway. The anti-angiogenic activity of osthole-PLGA-NPs/CS-FA was evaluated using the chick chorioallantoic membrane (CAM) assay. The results showed significant inhibition of angiogenesis in a concentration-dependent manner, starting at 40 μg/mL and increasing up to 120 μg/mL. In conclusion, osthole-PLGA-NPs/CS-FA nanoparticles exhibit promising potential for targeted pancreatic cancer therapy by enhancing cellular uptake, inducing apoptosis, and inhibiting angiogenesis.

Anticancer activity of Lannea coromandelica on B16F10 melanoma cell line: an in vitro and molecular docking approach

BackgroundPhytochemical screening was conducted on various bark extracts of Lannea coromandelica to assess their anticancer property against the B16F10 melanoma cell line. The phytoconstituents that were previously identified were utilized in molecular docking studies against the human tyrosinase related protein 1 (TYRP1) as a target receptor in order to provide more evidence for anticancer property. MethodsBark powder was extracted by maceration method using distilled water and soxhlet extraction using ethanol. The preliminary phytochemical evaluation and determination of total phenolic and flavonoid content of both extracts were conducted using biochemical assays. The present study investigated the possible anticancer effects of ethanol and aqueous extracts on the B16F10 melanoma cell line using the MTT assay and DAPI labelling techniques. The present study employed molecular docking techniques to assess the binding interactions between phytoconstituents and the TYRP1 protein, utilizing AutoDock Vina module of PyRx 0.8 software. ResultsThe phytochemical analysis found flavonoids, steroids, terpenoids, tannins, phenolic compounds, saponins, anthraquinones, cardiac glycosides, and proteins. Ethanolic extract shown preferential cytotoxicity to B16F0 melanoma cell line in-vitro (IC50= 9.69 ± 0.68 μg/ml), while aqueous extract exhibited IC50 = 75.49 ± 5.95 μg/ml. DAPI staining showed that treated cells had altered nucleus morphology, including apoptotic bodies. According to molecular docking investigations, Quercetin has the highest binding affinity (-9.6 Kcal/mol), followed by Catechin and Myricadiol. ConclusionThe current investigation has determined that Lannea coromandelica exhibits cytotoxic characteristic, as evidenced by the utilization of computer aided drug design models and in-vitro experimentation.

New Organoruthenium(II) Complexes Containing Andrographolide‐Appended Hydrazide Derivatives: Synthesis, Spectral Characterization, Anticancer Evaluation

ABSTRACTA new set of andrographolide‐appended hydrazide derivatives (AFC, AGC, and ATC) were prepared from the reaction of andrographolide with furan‐2‐carboxylic acid hydrazide (AFC), pyridine‐3‐carboxylic acid hydrazide (AGC), and thiophene 2‐carboxylic acid hydrazide (ATC), and their corresponding ruthenium(II) complexes (Ru‐AFC, Ru‐AGC, and Ru‐ATC) were synthesized by the direct reaction of [RuCl2(η6‐p‐cymene)]2 with ligands in dichloromethane under stirring condition. The compounds were analyzed by elemental analysis, IR, UV‐Vis, 1H NMR, and ESI‐MS spectrometric techniques, and the obtained spectral data revealed that the ligands coordinated to Ru(II) ion through their enone oxygen and amine nitrogen. Antioxidant activities of the ligands and complexes were calculated against DPPH•, ABTS•+, O2−, and NO• free radicals and their results were compared with standard antioxidants. The cytotoxic activity of the synthesized ligands and complexes toward A549 and HeLa cell lines was evaluated using MTT method (MTT = 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide). Among the compounds, the complex Ru‐AGC showed better cytotoxic effect against A549 and HeLa cancer cells with IC50 values of 4.75 ± 0.17 and 6.32 ± 0.26 μM, respectively. Further, the nontoxic nature of the compounds was confirmed by using human umbilical vein endothelial (HUVEC) cells. The apoptosis was inspected with AO/EB and DAPI staining methods; further, the percentages of the apoptotic and necrotic cells were determined by flow cytometry. Overall, the biological activities of our ruthenium(II)‐arene complexes were found to be more potent than their parent ligands due to chelation, and among the complexes, Ru‐AGC showed better activity due to the presence of pyridine ring in the N‐terminal position of the ligand. The obtained results highlighted the strong possibility to develop highly active ruthenium complexes as anticancer agents.

Anticancer effects of emetine against cisplatin resistant pancreatic cancer cells via activation of Cyto-c/Caspase-9/Caspase-3/Bax/Bad axis

This investigation examines the potential pharmacotherapeutic efficacy of emetine, a natural alkaloid, against pancreatic ductal carcinoma, utilizing the PaCa3 cell line as a model. Employing a multifaceted approach, the study explores emetine's impact on cell viability, clonogenic potential, and apoptotic pathways, unravelling its molecular mechanisms. Additionally, an in-vivo assessment is conducted to evaluate emetine's antitumor efficacy against PaCa3 xenografts. Under standard conditions, PaCa3 cells were treated with varying emetine concentrations and subjected to MTT, clonogenic, DAPI, AO/EB, Annexin V-FITC/PI, and Western blot assays. In-vivo experiments involved the administration of emetine to female nude mice with PaCa3 xenografts, with subsequent measurement of tumor weight and volume. Statistical analyses included Tukey-test and ANOVA. Emetine demonstrated a significant and dose- and time-dependent cytotoxic effect on PaCa3 cells, reducing viability from 98% to 8% (***p&lt;0.001). Clonogenic assay indicated a substantial inhibition of colony formation. DAPI staining revealed nuclear chromatin condensation, while AO/EB staining indicated heightened apoptosis. Annexin V-FITC/PI assay exhibited a dose-dependent decrease in normal cell viability and an increase in apoptotic events. Western blot analysis showcased concentration-dependent downregulation of anti-apoptotic Bcl-2 and upregulation of pro-apoptotic markers. In-vivo experiments illustrated a dose-dependent reduction in tumor weight (5 g to 1.6 g) and volume (2400 mm³ to 844 mm³) with emetine treatment. In conclusion, emetine emerges as a potent anticancer agent against pancreatic ductal carcinoma, exhibiting cytotoxicity, clonogenic inhibition, and apoptotic induction in PaCa3 cells. The in-vivo study further highlights its promising therapeutic potential, laying the groundwork for future clinical studies in pancreatic cancer treatment.

Antimicrobial cyclic lipopeptides from Bacillus mojovensis B1302 against wheat root rot

Bipolaris sorokiniana is the main pathogen affecting wheat, which can cause major disasters in wheat planting areas. In this study, cyclic lipopeptides (CLP) were extracted from Bacillus mojovensis B1302 and had antimicrobial activity on Bipolaris sorokiniana and other four pathogenic fungi. CLP could inhibit the growth of B. sorokiniana and made mycelia swelling and deformed. In addition, trypan blue staining indicated the distortion of mycelia and DAPI staining showed mycelial DNA damage in the CLP treatment group. Moreover, CLP were effective in controlling wheat root rot with the efficacy of 80.32%, which was not significantly different from that of commercial fungicide (carbendazim). Next, in the oil spreading test, the diffusion diameter of 40 μL CLP (10 mg/mL) reached 8.37 cm with a stable spreading ring, indicating CLP had surface activity. And also, CLP had strong antioxidant capacity and DPPH scavenging activity. Furthermore, CLP had better temperature tolerance at 20–40 °C and pH tolerance at 4–8, respectively, still maintained a high inhibition rate after 3h treatment under UV irradiation, and were stable under trypsin treatment. However, CLP were sensitive to pepsin and metal ions. In conclusion, CLP had antimicrobial activity, antioxidant capacity and environmental tolerance, which had good potential for development into biocontrol agents against wheat root rot.

In vitro anticancer efficacy of Calendula Officinalis extract-loaded chitosan nanoparticles against gastric and colon cancer cells

Objective This study assessed the anticancer activities of Calendula officinalis-loaded chitosan nanoparticles in gastric and colon cancer cells compared with fibroblast cells and examined the balance between ROS and antioxidants. Methods Considering this information, we synthesized Calendula officinalis-loaded chitosan nanoparticles (CO-CSNPs) via the ionic gelation method. Their characterizations were carried out with ZetaSizer, UV-Vis, FTIR and SEM devices including size, morphology and surface zeta potential analysis, loading capacity, encapsulation efficiency, in vitro drug release, and chemical interactions. The anticancer activities of CO, CSNPs, and CO-CSNPs were tested against AGS, Caco-2, and normal NIH-3T3 cells using an XTT assay. The anticancer effects were evaluated using DAPI staining, scratch assay, reactive oxygen species (ROS) detection and the CUPRAC method on cellular and non-cellular processes that promote anticancer mechanisms. Results Results showed that CO and CO-CNPs exhibited anticancer activity against AGS and Caco-2. Further, the formulation of CO with CSNPs enhanced the anticancer activity of CO while having no cytotoxicity on NIH-3T3. DAPI staining, scratch assay, ROS, and CUPRAC method confirmed the anticancer activity of CO and CO-CSNPs, which resulted in a reduction in the number of apoptotic cells, inhibited migration, triggered apoptotic pathway via ROS, and higher antioxidant activity. Conclusions The results of the study indicate that CO-CSNPs are a promising therapeutic formulation for gastric and colon cancer treatment. We consider that this study will lead to the investigation of molecular mechanisms of CO-CSNPs in cancer treatment and their investigation in clinical studies.

Co-crystals of thiobarbituric acid and N-heterocyclic compounds: Synthesis, structural, solubility, computational, and antilymphoma activity

Two new co-crystals (1–2) were prepared by the reaction of 2-thiobarbituric acid with N-heterocyclic compounds, and successfully characterized. X-ray analysis shows that 1 and 2 have zig-zag and ladder like supramolecular frameworks, respectively. HOMO-LUMO analysis of optimized geometries confirmed the lesser band gap of 1 and 2. Co-crystals 1–2 have solubility values of 19.737 and 25.652 mg/ml, respectively, and they may have good bioactivity. The cytotoxic activity of 1–2 indicated the tumor growth inhibitory ability of 1 and 2 against T cell lymphoma. Interestingly, apoptosis evaluation by Wright-Giemsa and DAPI staining suggested that 1 and 2 mediated tumor cell death through apoptosis induction. In addition, the estimation of glucose, pH, and NO also indicated the possible role of inhibited glucose uptake and augmented NO production in the cell death mediated by 1 and 2.

Baicalein induces apoptosis by targeting ribosomes in Candida auris.

The emergence of the "super fungus" Candida auris poses a significant threat to human health, given its multidrug resistance and high mortality rates. Therefore, developing a new antifungal strategy is necessary. Our previous research showed that Baicalein (BE), a key bioactive compound from the dried root of the perennial herb Scutellaria baicalensis Georgi, has strong fungistatic properties against C. auris. Nevertheless, the antifungal activity of BE against C. auris and its mechanism of action requires further investigation. In this study, we explored how BE affects this fungus using various techniques, including scanning electron microscopy (SEM), Annexin V-FITC apoptosis detection, CaspACE FITC-VAD-FMK In Situ Marker, reactive oxygen species (ROS) assay, singlet oxygen sensor green (SOSG) fluorescent probe, enhanced mitochondrial membrane potential (MMP) assay with JC-1, DAPI staining, TUNEL assay and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Our findings revealed that BE induced several apoptotic features, including phosphatidylserine (PS) externalization, metacaspase activation, nuclear condensation and DNA fragmentation. BE also increased intracellular ROS levels and altered mitochondrial functions. Additionally, transcriptomic analysis and RT-qPCR validation indicated that BE may induce apoptosis in C. auris by affecting ribosome-related pathways, suggesting that ribosomes could be new targets for antifungal agents, in addition to cell walls, membranes, and DNA. This study emphasizes the antifungal activity and mechanism of BE against C. auris, offering a promising treatment strategy for C. auris infection.

Betanin-encapsulated starch nanoparticles: synthesis and cytotoxic effect on colon cancer.

Colorectal cancer (CRC) is a common and life-threatening neoplastic disease that continues to pose a formidable challenge to global health. The present work was performed to evaluate the anticancer properties of betanin and betanin (BT) loaded starch nanoparticles (S-BT). The BT and S-BT were characterized by DLS, SEM, UV spectroscopy, XPS and FTIR. The cytotoxic effect was assessed by MTT and LDH assay. The apoptotic potential of BT and S-BT was assessed by DCFDA, Rh123, AO/EB and DAPI staining methods. Cell cycle arrest was depicted using flow cytometry. The antimetastatic potential of BT and S-BT was evaluated by wound healing assay. The S-BT showed a spherical morphology with a size of 175nm. The betanin contained SNPs were found to have strong encapsulation efficiency and favorable release profiles. Both BT and S-BT exhibited cytotoxicity in SW480 cells but S-BT displayed increased cytotoxicity when compared to BT alone. Loss of mitochondrial membrane potential, nuclear fragmentation, chromatin condensation and generation of ROS, all indicative of apoptotic mode of cell death, were revealed by fluorescence imaging. The cells were arrested in the G2M phase. Moreover, both BT and S-BT were able to inhibit the migratory potential of SW480 cells. Overall, our results indicated that both BT and S-BT were able to induce anticancer effects; and, S-BT was found to have increased therapeutic efficacy when compared to BT alone.

Investigating the anticancer potential and apoptotic signaling of nanocapsule-loaded pyrogallol in ovarian cancer cells (A2780)

Nanoencapsulation technology enables the efficient delivery of natural bioactive compounds, enhancing stability and controlled release to improve bioavailability and therapeutic efficacy. Pyrogallol, a natural polyphenolic compound, has shown promising anticancer activities against various cancer cell lines. The purpose of this study was to synthesize and characterize nanocapsule-encapsulated pyrogallol (Na-Pyr) and investigate its anticancer potential, apoptotic signaling, and anti-metastatic properties. The process involved synthesizing the nanocapsules using the sol–gel method and evaluating their physical and chemical properties using various techniques. The MTT assay was used to test Na-Pyr’s cytotoxicity against various cancer cell lines, including ovarian cancer (A2780), human colorectal adenocarcinoma (HT-29), and pancreatic cancer (ASPC-1), with Huvec serving as the normal cell line. DAPI staining, real-time PCR, and flow cytometry were performed to examine apoptosis induction and cell cycle arrest. The results obtained from dynamic light scattering (DLS) manifested that Na-Pyr had a size of 176.4 ± 7.26 nm, an average zeta potential of about 42.1 ± 2.01 mV, and a polydispersity (PDI) index of 0.174 ± 0.02. Electron microscopy images exhibited a smooth surface and spherical shape, indicating material compatibility. Na-Pyr exhibited higher anticancer potential against ovarian cancer cells than HT-29 and ASPC-1 cancer cell lines with an IC50 value of 23.5 ± 0.95 μg/mL. Na-Pyr significantly modulates the Bax/Bcl2 ratio and activates the MMP9 pathway-mediated apoptosis. Flow cytometry analysis supported these findings, revealing increased percentages of necrotic, early apoptotic, and late apoptotic populations upon exposure to Na-Pyr. In conclusion, the study highlights the promising anticancer potential of Na-Pyr and its potential as an effective treatment strategy.

Development of small-diameter vascular grafts using PCL/PLA/gelatin/PVA/hyaluronic acid nanofibers containing VEGF/enoxaparin

The extending demands for narrow-diameter vascular grafts to restore damaged arteries have captivated researcher's attention. The absence of appropriate vascular substitutes is due to the poor patency rate of small-caliber synthetic grafts caused by thrombosis and vessel hemodynamic mismatch that induce intimal hyperplasia. We developed small-diameter vascular grafts in two layers through electrospinning. The inner layer was fabricated by gelatin/polyvinyl alcohol (PVA)/hyaluronic acid/enoxaparin and vascular endothelial growth factor (VEGFs) and the outer layer was manufactured via polycaprolactone (PCL)/polylactic acid (PLA)/hyaluronic acid. The products were characterized by atomic force microscope (AFM), scanning electron microscope (SEM), water contact angle (WCA), attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), swelling test, mechanical test, and release rate. Blood compatibility tests including blood clotting time, hemolysis, and human platelet sedimentation were measured. The interactions of human umbilical vein endothelial cells (HUVECs) and human endometrial stem cells (hEnSCs) with nanofibrous vascular grafts were detected by SEM, viability assay, and DAPI staining. The results exhibited the produced small-diameter vascular grafts had proper antithrombotic properties due to two factors of VEGF and enoxaparin. After two months of the subcutaneous implantation of the scaffolds on the back of rats, the tubular vascular grafts lacked any macroscopic and histological signs of acute inflammation and had appropriate degradation rates. Furthermore, lumen patency was observed after 2 months. Consequently, VEGF and enoxaparin improves cell and blood compatibility in the small-diameter vascular grafts.

CYP1B1 promotes PARPi-resistance via histone H1.4 interaction and increased chromatin accessibility in ovarian cancer

IntroductionOvarian cancer is the most lethal gynecological cancer and presents significant therapeutic challenges. The discovery of synthetic lethality between PARP inhibitors (PARPi) and homologous recombination deficiency marked a new era in treating BRCA1/2-mutated tumors. However, PARPi resistance remains a major clinical challenge. MethodsRNA sequencing was used to identify genes altered by PARPi treatment and LC-MS was used to detect proteins interacting with CYP1B1. Resistance mechanisms were explored through ATAC-seq and gene expression manipulation. Additional techniques, including micrococcal nuclease digestion assays, DAPI staining, and fluorescence microscopy, were used to assess changes in nuclear morphology and chromatin accessibility. ResultsThe gradual exposure of Olaparib has developed a PARPi-resistant cell line, A2780-OlaR, which exhibits significant upregulation of CYP1B1 at both RNA and protein levels. Down-regulating CYP1B1 expression or using specific inhibitors decreased the cellular response to Olaparib. Linker histone H1.4 was identified as associated with CYP1B1. ATAC-seq showed differential chromatin accessibility between A2780-OlaR and parental cells, indicating that the downregulation of H1.4 was associated with increased chromatin accessibility and higher cell viability after Olaparib treatment. ConclusionOur findings reveal a novel role for CYP1B1 in driving PARPi resistance through distinct molecular mechanisms in A2780-OlaR. This study highlights the importance of chromatin accessibility in PARPi efficacy and suggests the CYP1B1/H1.4 axis as a promising therapeutic target for overcoming drug resistance in ovarian cancer, offering potentially therapeutic benefits.

Effects of different slaughtering methods on the energy metabolism, apoptosis process and quality of grouper (Epinephelus fuscoguttatus) during cold storage at 4 °C.

The aquatic processing industry is increasingly aware of the need to ensure that slaughtering is carried out under high welfare standards, so there is a need to explore the impact of slaughter methods on fish fillets. This study aimed to investigate the effects of different slaughtering methods (M1, lethality by hammering; M2, gas mixture causing death; M3, lethality by clove oil anesthesia + ice slurry; M4, lethality by ice slurry; M5, lethality by gradient cooling) on the energy metabolism, apoptosis and flesh mass in grouper (Epinephelus fuscoguttatus). Therefore, 120 fish (24 per treatment) were slaughtered by the five methods. The results showed that the succinate dehydrogenase (SDH) enzyme activity of M5 sample was higher. The serum glucose level of M2 samples and DAPI staining fluorescence of M2 samples were the highest, indicating that the stress response of M2 was strong. In addition, the texture, pH, total volatile basic nitrogen (TVB-N), thiobarbituric acid (TBA) and K value results showed M5 samples had better flesh quality. Gradient cooling lethality had the least effect on oxidative damage and apoptosis in grouper during cold storage as the gradient cooling lethality had the least effect on antioxidant enzyme activities. © 2024 Society of Chemical Industry.

Morin promotes autophagy in human PC3 prostate cancer cells by modulating AMPK/mTOR/ULK1 signaling pathway

AMP-activated protein kinase (AMPK) suppresses tumorigenesis by modulating autophagy and apoptosis. This study evaluated the impact of Morin on PC3 prostate cancerous cells by examining the AMPK/ mechanistic target of rapamycin (mTOR)/ ULK1 (UNC-51-like kinase 1) pathway and autophagy process. The PC3 cells were treated with Morin (50 µg/ml) and AICAR (an AMPK activator). Cell viability, apoptosis, autophagy, and level of phosphorylated and non-phosphorylated ULK1, AMPK, and mTOR, as well as LC3B/LC3A, have been investigated. Through DAPI staining, measurement of Bax/Bcl-2 ratio, Caspase activity, and Annexin V/PI method, it has been revealed that Morin induces apoptosis and reduces the growth of PC3 cells. Morin enhanced the protein level of phosphorylated AMPK (p-AMPK) and ULK1 (p-ULK1) and decreased the expression of phosphorylated mTOR (p-mTOR) in the PC3 cells. Morin could also increase the LC3B/LC3A ratio, Acridine Orange-positive cells, expression of Beclin-1 and ATG5 genes, and decrease the p62 protein level indicating autophagy-inducing. AICAR (an AMPK activator) enhanced the impact of Morin on apoptosis, cell growth, and expression of LC3B, p-AMPK, p-ULK1, and p-mTOR proteins in the PC3 cells. These findings suggest that Morin induces apoptotic and autophagic cell death by activating AMPK and ULK1 and suppressing mTOR pathways.