Lower IC50 Research Articles

Phytochemical investigation and evaluation of antioxidant and antidiabetic activities in aqueous extracts of Cedrus atlantica

Abstract Diabetes mellitus is a global health problem requiring innovative approaches for effective management. Natural compounds derived from medicinal plants offer promising avenues due to their diverse biological activities. The aim of this study was to assess the inhibitory effects of Cedrus atlantica extracts on the enzymes α-amylase and α-glucosidase, which play a crucial role in the regulation of postprandial glucose levels, as well as to investigate their phytochemical composition and antioxidant capabilities. Aqueous extract of bark (EA), aqueous extract of cones (CA), and aqueous extract of leaves (FA) of C. atlántica were prepared and evaluated for enzyme inhibition using acarbose as a standard. The CA extract showed potent α-amylase inhibition (IC50 307 ± 0.02 μg/mL) and remarkable α-glucosidase inhibition (IC50 70 ± 0.04 μg/mL), outperforming the FA and EA extracts. Quantitative analysis revealed that EA and CA extracts had higher total phenolic content, flavonoid content, and tannin content than FA extract. Antioxidant assessments highlighted the outstanding performance of CA extract, with a low IC50 ABTS (30.65 ± 0.1 μg/mL) and an impressive EC50 FRAP (59.43 ± 1.19 μg/mL), outperforming FA extract. The results demonstrate the remarkable antidiabetic potential of C. atlantica extracts, particularly the CA extract, by inhibiting key enzymes involved in carbohydrate digestion. These extracts possess various phytochemical compounds with significant antioxidant capacities, suggesting that they are suitable for pharmaceutical, nutraceutical, and cosmetic applications. Further research to isolate and identify the bioactive compounds in CA extract could lead to new therapeutic strategies for managing diabetes and oxidative stress-related conditions.

Apoptotic cell death of stomach cancer lines (AGS) induced by Co-NTB complex through cellular organelles and DNA damage.

Given that stomach cancer is the fourth leading cause of cancer-related death, there is a need to develop new drugs. Among various methods, metal-based coordination compounds are considered as an efficient strategy against this type of cancer. Similarly, the benzimidazole moiety plays a crucial role in biology; thus, various benzimidazole-based compounds have been found to be active as potential anticancer drugs and are currently used in clinical trials. In this study, we explored the benzimidazole-based cobalt(ii) complex as an anticancer agent against AGS stomach cancer cell lines. Interestingly, the MTT assay of the Co-NTB complex shows a lower IC50 value of 4.25 μg mL-1 compared to cisplatin, which has an IC50 of 7.5 μg mL-1 against AGS cell lines. Light microscopy and Hoechst/propidium iodide dye staining clearly indicate that the complex damages DNA, leading to cell death through an apoptotic pathway. The apoptotic cell death pathway was further complemented by Lysotracker and Mitotracker staining, as well as transmission electron microscopy (TEM) imaging. Overall, the Co-NTB complex acts as an effective anticancer agent against AGS stomach cancer cell lines, with apoptotic cell death induced by targeting cellular organelles and DNA.

Construction and validation of prognosis and treatment outcome models based on plasma membrane tension characteristics in bladder cancer.

Plasma membrane tension-related genes (MTRGs) are known to play a crucial role in tumor progression by influencing cell migration and adhesion. However, their specific mechanisms in bladder cancer (BLCA) remain unclear. Transcriptomic, clinical and mutation data from BLCA patients were collected from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Clusters associated with MTRGs were identified by consensus unsupervised cluster analysis. The genes of different clusters were analyzed by GO and KEGG gene enrichment analysis. Differentially expressed genes (DEGs) were screened from different clusters. Consensus cluster analysis of prognostic DEGs was performed to identify gene subtypes. Patients were then randomly divided into training and validation groups, and MTRG scores were constructed by logistic minimum absolute contraction and selection operator (LASSO) and Cox regression analysis. We assessed changes in clinical outcomes and immune-related factors between different patient groups. The single-cell RNA sequencing (scRNA-seq) dataset for BLCA was collected and analyzed from the Tumor Immune Single-cell Hub (TISCH) database. Biological functions were investigated using a series of experiments including quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), wound healing, transwell, etc. Our MTRG score is based on eight genes (HTRA1, GOLT1A, DCBLD2, UGT1A1, FOSL1, DSC2, IGFBP3 and TAC3). Higher scores were characterized by lower cancer stem cell (CSC) indices, as well as higher tumor microenvironment (TME) stromal and immune scores, suggesting that high scores were associated with poorer prognosis. In addition, some drugs such as cisplatin, paclitaxel, doxorubicin, and docetaxel exhibited lower IC50 values in the high MTRG score group. Functional experiments have demonstrated that downregulation of DCBLD2 affects tumor cell migration, but not proliferation. Our study sheds light on the prognostic significance of MTRGs within the TME and their correlation with immune infiltration patterns, ultimately impacting patient survival in BLCA. Notably, our findings highlight DCBLD2 as a promising candidate for targeted therapeutic interventions in the clinical management of BLCA.

Deciphering physical and functional properties of chitosan-based particles for agriculture applications

Traditional methods for controlling plant pathogens rely on toxic chemicals, posing environmental and health risks. Developing sustainable, eco-friendly alternatives is crucial. Chitosan (CS)-based materials offer promising solutions for sustainable agriculture. We aimed to synthesize and characterize CS-based microparticles with varying properties and assess their antimicrobial performance to establish correlations between variations in physicochemical characteristics and their impact on performance within biological systems. We adjusted the synthesis parameters, producing particles labeled P1, P2, and P3, which have sizes of 0.19 ± 0.07 μm, 0.45 ± 0.32 μm, and 1.22 ± 0.32 μm, and zeta potentials of +7.6 ± 4.25 mV, +22 ± 3.51 mV, and + 12.9 ± 4.54 mV, respectively. Extensive toxicological screenings showed that these CS-based microparticles were non-toxic across cell cultures, mouse red blood cells, soil microbiota, nitrogen-cycling bacteria, and plant toxicity assays. Encouraged by these results, we evaluated their antimicrobial potential against economically important crop pathogens. The CS-based microparticles exhibited antimicrobial effects against the bacterium Pseudomonas syringae pv. tomato DC3000 and the fungus Fusarium solani f. sp. eumartii. Higher zeta potentials correlated with greater antimicrobial efficacy, evidenced by lower IC50 and minimum inhibitory concentration (MIC) values. These findings indicate that all three microparticles analyzed displayed antimicrobial activity against two economically significant crop pathogens, with P2 showing solid performance attributed to its physicochemical characteristics. Therefore, CS-based microparticles represent a promising, nontoxic, and environmentally friendly alternative for modern agriculture, with their biological activities potentially predictable through careful selection of physicochemical properties before the synthesis process.

Anti-inflammatory 5,6,7,8-tetrahydro-2-(2-phenylethyl) chromone derivatives from the stems of Aquilaria sinensis

Three previously undescribed 5,6,7,8-tetrahydro-2-(2-phenylethyl)chromones, (5S,6R,7S,8S)-8-chloro-5-ethoxy-6,7-dihydroxy-2-[2-(4′-methoxyphenyl)ethyl]-5,6,7,8-tetrahydrochromone (1), (5R,6S,7R,8R)-8-chloro-5-ethoxy-6,7-dihydroxy-2-(2- phenylethyl)-5,6,7,8-tetrahydrochromone (2), (5S,6R,7S,8R)-5,8-dichloro-6,7- dihydroxy-2-[2-(4′-methoxyphenyl)ethyl]-5,6,7,8-tetrahydrochromone (3), and 28 known analogues (4–31) were isolated from the stems of Aquilaria sinensis. Their structures were characterized by the spectroscopic methods, and the absolute configuration was resolved by circular dichroism (CD) spectroscopy. Bioactivity evaluation indicated that compounds 3–8 had significant inhibition effect in the production of NO in an inflammatory cell model with relatively lower IC50 values of 5.54, 11.44, 3.68, 7.15, 10.26 and 13.04 μM, respectively, compared to the positive control Indomethacin (IC50 = 23.03 μM).

Investigating the P53-dependent anti-cancer effect of ibutamoren in human cancer cell lines.

The MDM2-p53 pathway plays a pivotal role in regulating cell cycle and apoptosis, with its dysfunction contributing to approximately 50% of human malignancies. MDM2, an E3 ubiquitin ligase, targets the tumour suppressor p53 for degradation, thereby promoting uncontrolled cell growth in cancers. Inhibiting the MDM2-p53 interaction represents a promising therapeutic strategy for reactivating p53's tumour-suppressive functions. This study explored the potential of ibutamoren (IBU) as a novel inhibitor of MDM2. In silico analyses utilizing molecular modelling revealed that IBU has a low IC50 for MDM2 inhibition and favourably binds to the p53-binding pocket of MDM2. In vitro experiments demonstrated that IBU treatment reduced the viability of immortalized cancer cell lines with a functional MDM2-p53 pathway but not in cell lines where this pathway harboured damaging mutations. This trend was further supported by RT-qPCR analysis, which showed differential expression of two p53 target genes upon IBU treatment in cell lines with wild MDM2-p53 pathways but not in those harbouring damaging mutations. These findings provide preliminary evidence supporting IBU's anticancer activity, plausibly through the MDM2-p53 pathway, and suggest that further studies are warranted to explore its mechanism of action and potential development as a lead compound in oncology research.

Development, Optimization, Characterization, and In-vitro Assessment of PEG-coated Docetaxel-loaded Proliposomes for Delivery to Breast Cancer Cells

Introduction: To address the issues of liposomal instability and enhance the delivery of docetaxel (DTX) to breast cancer cells, the development of polyethylene glycol (PEG)-coated proliposomes was proposed, utilizing a thin film hydration technique followed by lyophilization with an appropriate cryoprotectant. Various compositions of phospholipid, cholesterol, and docetaxel were optimized, and the PEG was used in a 1.3:1 ratio of the phospholipid. Method: The liposomes were converted to proliposomes using a lyophilizer. The optimized formulation possesses a particle size of 117.0 ± 9.78 nm, with a polydispersity index (PDI) of 0.265 ± 0.094, drug entrapment (DE) of 96.0± 6.14 %, and drug loading (DL) of 9.20 ± 3.17 %. In-vitro study demonstrated a controlled release pattern consistent with the Higuchi model, alongside significantly lower protein binding relative to free DTX, indicating a potential reduction in side effects. Results: Cell viability study demonstrated increased cytotoxicity of PEG-DTX proliposomes (PEGDTX PL) against MDA-MB-231 cells, evidenced by a lower IC50 relative to free DTX, underscoring the promise of this nanocarrier for targeted therapy. We found IC50 values of 11.885 μg/mL and 4.677 μg/mL for DTX and PEG-DTX PL (p<0.05). Conclusion: The findings are promising as a simple and scalable carrier comprising general and biocompatible materials that can provide a safe surfactant free nanosystem with improved efficacy and performance.

Assessing the Impact of the Leader Peptide in Protease Inhibition by the Microviridin Family of RiPPs.

Background: Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a growing class of natural products biosynthesized from a genetically encoded precursor peptide. RiPPs have attracted attention for the ability to generate and screen libraries of these compounds for useful biological activities. To facilitate this screening, it is useful to be able to do so with the leader peptide still present. We assessed the suitability of the microviridin family for these screening experiments by determining their activity with the leader peptide still present. Methods: Modified precursor peptides with the leader present were heterologously expressed in Escherichia coli. Their ability to inhibit elastase was tested with a fluorogenic substrate. HPLC was used to monitor degradation of the modified precursor peptides by elastase. SDS-PAGE was used to determine the ability of immobilized modified precursor peptide to pull down elastase. Results: We found that the fully modified precursor peptide of microviridin B can inhibit the serine protease elastase with a low nanomolar IC50, and that the fully modified precursor with an N-terminal His-tag can mediate interactions between elastase and Ni-NTA resin, all indicating leader peptide removal is not necessary for microviridins to bind their target proteases. Additionally, we found that a bicyclic variant was able to inhibit elastase with the leader peptide still present, although with a roughly 100-fold higher IC50 and being subject to hydrolysis by elastase. Conclusions: These results open a pathway to screening libraries of microviridin variants for improved protease inhibition or other characteristics that can serve as, or as inspirations for, new pharmaceuticals.

Phytofabrication of silver nanoparticles by using Cucurbita maxima leaf extract and its potential anticancer activity and pesticide degradation

ABSTRACT Global cancer mortality is rising yearly due to various factors. Silver nanoparticles (AgNPs) are widely utilized for their low cost and antibacterial properties compared to other noble metals. Here, AgNPs were synthesized using Cucurbita maxima leaf extract, which was analyzed by various analytical instruments. UV-visible spectra showed an absorbance peak in the 410-420 nm range, while the Fourier transform infrared exhibited bands at 653 cm−1, 1629, 2276, and 3786 cm−1. X-ray diffraction showed major intensity peaks at 38.1º two theta value with an average crystallite size of 18.34 nm. The transmission electron microscope showed the aggregation of the spherical-shaped AgNPs, the size of which was 10-90 nm. The MTT assay of the AgNPs showed that it is highly effective against SiHa cervical cancer cells with a much lower IC50 value (~8 µg/mL) and capable of pesticide removal with the highest efficiency at pH 9.

In vitro Antioxidant and Antitumor Activity of Rooibos (Aspalathus linearis) Extract on Colon, Prostate, and Liver Cancer Cells

Rooibos tea (Aspalathus linearis) is recognized for its antioxidant properties, due to distinctive compounds such as aspalathin. This study examined the antioxidant potential of rooibos extract on three cancer cell lines (PC-3, HCT-116, and HepG2) to assess its overall antioxidant potential. Additionally, the inhibitory effect of aqueous rooibos extract at a concentration of 1,000 µg/ml was assessed on the three cancer cell lines. The cytotoxic effects of aqueous rooibos extract and cisplatin were compared for the cancer cell lines, with HepG2 cells showing the highest sensitivity to rooibos extract (IC50 = 1,399.41 ± 62.73 µg/ml) and PC-3 cells displaying the highest resistance (IC50 = 2,431.51 ± 98.71 µg/ml). Cisplatin consistently exhibited lower IC50 values across all cell lines, indicating a stronger inhibitory effect on cancer cell proliferation, notably for HCT-116 (IC50 = 2.53 ± 0.21 µg/ml) and HepG2 cells (IC50 = 3.67 ± 0.25 µg/ml). However, the total antioxidant capacity of rooibos extract (68.91 ± 4.07 mg GAE/g) is comparable to that of the ascorbic acid standard (74.19 ± 3.97 mg GAE/g), showing no statistically significant difference. Moreover, rooibos extract is a potent antioxidant, with lower IC50 values in the DPPH (3.62 ± 0.10 µg/ml), ABTS (2.13 ± 0.13 µg/ml), and H2O2 assays (4.97 ± 0.01 µg/ml) compared to ascorbic acid and EDTA. However, in the metal chelating activity assay, rooibos extract showed a similar performance (IC50 20.89 ± 0.09 µg/ml) to EDTA (IC50 21.16 ± 1.67 µg/ml), suggesting comparable efficacy in this measure. These results demonstrate the strong antioxidant and anticancer potential of rooibos extract, suggesting it as a promising complement in cancer treatment.

Construction of Cisplatin-18-Crown-6 Complexes Through Supramolecular Chemistry to Improve Solubility, Stability, and Antitumor Activity.

Cisplatin (DDP), a platinum-chelated compound renowned for its antitumor activity, is often utilized in cancer therapy. However, its real-world clinical efficacy is compromised by poor solubility and low stability, which impedes wider clinical application. Our study aimed to address these limitations of DDP through host-guest supramolecular chemistry approaches. We explored the potential of 18-crown-6 as the host molecule to solubilize and stabilize DDP, the guest molecule. Utilizing techniques such as UV-visible spectroscopy, Fourier-transform infrared spectroscopy, Raman spectroscopy, and molecular docking, we conducted a comprehensive analysis on the physical state and inclusion mode of the DDP@18-crown-6 complex. Phase solubility studies and Job's plot confirmed that the DDP@18-crown-6 complex significantly enhanced the aqueous solubility of DDP, with an optimal 1:1 binding ratio. Stability analyses revealed that this complex markedly improved the stability of DDP in pure water. Meanwhile, the stabilization effects of DDP@18-crown-6 were remarkably elevated when combined with 0.9% sodium chloride. In vitro antitumor assays in A549 cell lines demonstrated that the DDP@18-crown-6 complex outperformed raw DDP in cytotoxicity, showing a significantly lower IC50 value. This research offered a promising strategy for DDP solubilization and stabilization, facilitating its anticancer therapeutic efficacy.

Rosthornin B alleviates inflammatory diseases via directly targeting NLRP3.

Aberrant activation of the NLRP3 inflammasome contributes to the evolution of diverse inflammatory diseases. Inhibition of the NLRP3 inflammasome has been proven to be an effective treatment strategy for NLRP3-driven diseases. This study revealed that multiple natural diterpenes from Isodon plants can inhibit the NLRP3 inflammasome, among which Rosthornin B (Ros B) exhibited the best inhibitory effect, with an IC50 of 0.39 μM. Further study revealed that Ros B directly interacts with NLRP3, thereby restraining NEK7-NLRP3 interaction and inhibiting NLRP3 inflammasome assembly and activation. Remarkably, Ros B had a significant therapeutic benefit in mouse models of NLRP3-driven septic shock, peritonitis, and colitis. Our study has identified a series of natural diterpenes that target the NLRP3 inflammasome. These natural diterpenes, especially those with low IC50 values, may lead to the development of new drugs and potential clinical therapies for diseases driven by NLRP3 inflammasome activation.

Discovery of a Novel Co-crystal of Chrysin and Oroxylin a with Anticancer Properties from Leaves of Oroxylum indicum.

As the number of new cancer cases increases every year, there is a necessity to develop new drugs for the treatment of different types of cancers. Plants' resources are considered to be huge reservoirs for therapeutic agents in nature. Among all the medicinal plants, Oroxylum indicum is one of the most widely used medicinal plants in India, China, and Southeast Asian countries. Combinatorial drug treatment, on the other hand, is favored over single drug treatment in order to target multiple biomolecular moieties that help in the growth and development of cancer. Therefore, combinatorial drug treatment using a co-crystal of multiple drugs gives researchers an idea of the development of a new type of drug for targeting multiple targets. In this study, a new co-crystal of chrysin and oroxylin A was isolated from the leaves of O. indicum, and its anticancer properties were studied in cervical cancer cells HeLa. This study was conducted with the aim of identifying new anticancer compounds from the leaves of Oroxylum indicum and studying the anticancer properties of the isolated compound. In this study, we elucidated the structure of a new co-crystal compound, which was isolated from the leaf extract of Oroxylum indicum. The apoptosis induction mechanism of the newly discovered co-crystal in HeLa cells was also studied. A crystal compound from the chloroform extract of leaves of Oroxylum Indicum was isolated by solvent fractionation and chromatographic methods involving HPLC. The molecular structure of the isolated crystal was elucidated by Single Crystal-XRD, FT-IR analysis, and further determined by LC-MS. The antiproliferative activity was carried out using an MTT assay and fluorescence microscopy, and the mechanism of apoptosis was determined using Western blotting techniques. The novel co-crystal consists of two active pharmaceutical ingredients (APIs) in a 1:1 ratio, i.e., oroxylin A and chrysin. The isolated new co-crystal induced death in HeLa cells with a very low IC50 value of 8.49μM. It induced caspase-dependent apoptosis in HeLa cells by activation of Caspase-3 through inhibition of ERKs and activation of p38 of MAPK cell signalling pathway. This study presents the first report on the discovery of a naturally occurring co-crystal of chrysin and oroxylin A and the involvement of ERKs and p38 of MAPK pathways in the induction of apoptosis in HeLa cells by the co-crystal. Our study sheds light on the development of a co-crystal of chrysin and oroxylin A in a specific ratio of 1:1 for combination therapy of the two APIs. The purified co-crystal was found to be more efficient compared to the compounds present individually. Further analysis of the physiochemical properties and molecular mechanisms of the isolated co-crystal in different cancer cells is warranted for its application in therapeutics.

Semisynthesis and Antitumour Evaluation of Natural Derivatives from ent-Kaurene ent-15α-Angeloyloxykaur-l6-en-3β-ol Isolated from Distichoselinum tenuifolium.

Two natural ent-kaurene diterpenoids, ent-15α-angeloyloxykaur-16-en-3β-ol (7) and ent-15α-angeloyloxykaur-16-en-3β,9-diol (8), were extracted from the aerial parts of Distichoselinum tenuifolium, and six new derivatives were synthesised from compound (7). The antitumour properties of these natural and derivative ent-kaurenes (2, 7, 9-13) were evaluated in three cancer cell lines: HT29 (colon cancer), HepG2 (hepatocellular carcinoma), and B16-F10 (murine melanoma). Among them, the synthesised ent-kaurene (13) containing an exomethylene-cyclopentanone moiety showed the strongest antiproliferative effects in all cell lines tested, with significantly lower IC50 values around 2.5 μM. Compounds 13 and 12, together with their precursor (7), were selected for further comparative cytometric and microscopic analyses. Cell cycle studies revealed that derivatives 12 and 13 exhibited promising cytostatic activity by inducing selective G2/M phase arrest, particularly effective in HT29 and HepG2 cells. Conversely, precursor (7) showed no significant effect on B16-F10 cell cycle distribution. The Annexin V-FITC/PI double staining assay confirmed the robust apoptotic effects of compounds (7), 12 and 13, with compound 13 inducing up to 99% total apoptosis and exhibiting significant apoptotic activity in all cell lines tested. These apoptotic effects were closely linked to mitochondrial dysfunction, as evidenced by a marked loss of mitochondrial membrane potential and reduced Rh123 fluorescence in treated cells, thereby activating the intrinsic apoptotic pathway. These findings highlight the critical role of mitochondrial disruption in the cytotoxic mechanisms of these ent-kaurenes and underscore their potential as promising anticancer agents.

Formulation of Biological Sunscreen from Calendula arvensis Capitula Extracts: Antioxidant, Anti-Aging, Surface Tension, and UVB Protection Properties Assessed

Skin protection against ultraviolet (UV) radiation has long been crucial due to its role in photoaging, sunburn, and wrinkles. This study focuses on developing a bio-based sunscreen from Calendula arvensis capitula extract. Various extraction methods (maceration, sonication, and infusion) and solvents (EtOH, EtOH-H2O, and H2O) were explored in order to identify the most effective extract for use in a sunscreen formulation. Each extract was analyzed for its phenolic content, as well as antioxidant activities (assessed through DPPH, CAT, and FRAP assays). Enzyme inhibition assays for tyrosinase, elastase, and collagenase highlighted the low IC50 values of the hydroethanolic extract. Furthermore, the in vitro sun protection factor (SPF) against UVB radiation was measured using ultraviolet spectrophotometry. A phytochemical analysis showed phenolic levels between 8 and 27 mg GAE/g, flavonoid concentrations of 7–13 mg QE/g, and tannin levels of 1.15–1.68 mg/mL, alongside moderate antioxidant activity. The ethanol maceration extract reduced the interfacial tension to 2.15 mN/m in 600 s, outperforming the conventional emulsifier polysorbate 20. The sonicated hydroethanolic extract demonstrated remarkable SPF efficacy (SPF = 193.65 ± 0.02), far exceeding that of the standard zinc oxide (SPF = 11.88 ± 0.03). The proposed formulations meet the COSMOS standards, suggesting their potential for certification as biological products. Further clinical and in vivo studies are necessary to confirm their safety and commercial viability.

Establishment and characterization of NCC-GCTB10-C1: a novel cell line derived from a patient with recurrent giant cell tumor of bone.

Giant cell tumor of bone (GCTB) is a rare osteolytic tumor composed of mononuclear stromal cells, macrophages, and osteoclast-like giant cells. While generally benign, GCTB has a high risk of local recurrence and can occasionally undergo malignant transformation or metastasis, posing significant clinical challenges. The primary treatment is complete surgical resection; however, effective management strategies for recurrent or advanced GCTB remain elusive, underscoring the need for further preclinical research. This study reports the establishment of a novel cell line, NCC-GCTB10-C1, derived from a recurrent GCTB lesion. NCC-GCTB10-C1 retains the characteristic H3-3A G34W mutation, which is central to the tumor's pathogenesis, and demonstrates significant growth potential, spheroid formation capability, and invasive properties. Extensive drug screening of NCC-GCTB10-C1, along with nine previously established GCTB cell lines, revealed a distinct drug response profile, with the cell line showing resistance to many previously effective agents. However, doxorubicin, foretinib, and ceritinib were identified as promising therapeutic candidates due to their low IC50 values in NCC-GCTB10-C1. The establishment of NCC-GCTB10-C1 offers a critical resource for further research into GCTB, especially in the context of recurrent disease, and holds potential for the development of more effective treatment strategies.

Establishment and characterization of a novel patient-derived cell line from conventional central grade 3 chondrosarcoma, NCC-CS1-C1.

Chondrosarcoma (CS) is a malignant tumor that produces cartilaginous matrix and is the second most common primary bone sarcoma. CS encompasses a range of histological subtypes, with high-grade conventional central CS being particularly rare, occurring at a rate of 1.81 cases per 1 million person-years. Complete surgical resection is the standard curative treatment for this subtype, as radiation therapy and chemotherapy have proven ineffective. High-grade conventional central CS is highly metastatic and prone to recurrence, resulting in a poor prognosis. Therefore, effective multidisciplinary treatment strategies are urgently needed. Patient-derived cell lines offer promising tools for exploring new therapeutic approaches. However, only two cell lines of high-grade CSs are currently available in public cell banks. In this study, we aimed to establish a novel cell line for high-grade conventional central CS. We successfully developed the NCC-CS1-C1 cell line using surgically resected tumor tissues from a patient with conventional central grade 3 CS. This cell line harbored an IDH1 mutation (p.R132S), commonly found in 50% of CS cases, and exhibited complex copy number variants. A high-throughput screening of 221 anti-cancer drugs identified five candidates-bortezomib, carfilzomib, doxorubicin, panobinostat, and romidepsin-that demonstrated low IC50 values, indicating potential efficacy in treating CS. These findings suggest that NCC-CS1-C1 is a valuable tool for both preclinical and basic research on high-grade conventional central CS.

Phenolic Compounds, Antioxidant Activity, Sensorial Analysis and Metabolic Syndrome Enzyme Inhibition Properties of Calafate (Berberis microphylla) Fruit Juice Fermented by Patagonian Lactic Acid Bacteria.

Calafate is a native barberry of Patagonia, used in culinary and medicinal practices since ancient times. The aim of this work was to analyze the ability of lactic acid bacteria (LAB) isolated from calafate fruits and flowers, to increase the phenolic compound concentration and antioxidant capacity as well as to inhibit metabolic-related enzymes in fermented calafate juices. The sensory attributes of the selected fermented juice were also analyzed. The LAB strains grew between 1.33 ± 0.03 and 2.61 ± 0.30 log CFU/ml in the calafate juices at 24h. Fructobacillus fructosus B7 consumed glucose and fructose the most (2.30 ± 0.45g/L and 3.73 ± 0.44g/L, respectively) and produced mannitol (3.89 ± 0.77g/L). The fermented juice by Lacticaseibacillus paracasei B4 showed the highest total phenolic compound concentration (2662.58 ± 344.51mg GAE/100ml) and antioxidant capacity (38916.42 ± 2157.52 µmol TE/100ml). The fermented juices inhibited the activity of metabolic syndrome-related enzymes. The lower IC50 for α-glucosidase activity was observed for F. fructosus B7 and L. paracasei B4 (0.56 ± 0.10 and 0.64 ± 0.05mg GAE/ml, respectively) fermented juices, while for α-amylase the lowest IC50 values were determined for the L. curvatus B34 and L. paracasei B4 (0.34 ± 0.01 and 0.37 ± 0.06mg GAE/ml, respectively) juices. The relative amount of isorhamnetins, which can induce GLUT4 translocation to the plasma membrane preventing hyperglycemia, was increased in the L. paracasei B4 fermented juice. The L. paracasei B4 fermented juice had acceptable sensorial values for consumption.

Novel active Trp- and Arg-rich antimicrobial peptides with high solubility and low red blood cell toxicity designed using machine learning tools.

Given the rising number of multidrug-resistant (MDR) bacteria, there is a need to design synthetic antimicrobial peptides (AMPs) that are highly active, non-hemolytic, and highly soluble to act as alternatives to antibiotics that are either no longer effective or used as drugs of last resort. Machine learning tools allow the straightforward in silico identification of non-hemolytic antimicrobial peptides. Here, we utilized a number of these tools to rank the best peptides from two libraries: 1) 8192 peptides with sequence bhxxbhbGAL, where b is the basic amino acid R or K, h is a hydrophobic amino acid, i.e. G, A, L, F, I, V, Y, or W and x is Q, S, A, or V; and 2) 512 peptides with sequence RWhxbhRGWL, where b and h are as for the first library and x is Q, S, A, or G. The top 100 sequences from each library, as well as 10 peptides predicted to be active but hemolytic (for a total of 220 peptides), were SPOT synthesized and their IC50 values were determined against S. aureus USA 300 (MRSA). Of these, 6 AMPs with low IC50's were characterized further in terms of: MICs against MRSA, E. faecalis, K. pneumoniae, E.coli and P. aeruginosa; RBC lysis; secondary structure in mammalian and bacterial model membranes; and activity against cancer cell lines HepG2, CHO, and PC-3. Overall, the approach yielded a large family of active antimicrobial peptides with high solubility and low red blood cell toxicity. It also provides a framework for future designs and improved machine learning tools.

Co-culture system of breast cancer and normal cells to investigate inflammation: using doxorubicin encapsulated in adipose-derived exosomes.

Doxorubicin (DOX) chemotherapy for breast cancer is an effective treatment option, but it also has disadvantages. Exosomes (EXOs) have safely and successfully transported DOX and reduced its adverse effects; however, its use is still being explored. In this study, a co-culture system of malignant and non-malignant breast cells was used to generate an in vitro model reflecting the in vivo cellular microenvironment, and the effects of this treatment were investigated by examining inflammatory genes. Extracellular matrices (EXOs) were extracted from mesenchymal stem cells derived from human adipose tissue by ultracentrifugation. Later, Western blotting, dynamic light scattering (DLS) and transmission electron microscopy methods were used to examine the properties of the EXO. DOX was encapsulated in the EXOs by sonication and the loading rate was measured by spectrophotometry. In the current study, a co-culture system was used to investigate the cytotoxic effects of free DOX and DOX encapsulated in EXOs (EXO-DOX) on various breast cell lines, including MCF-7, MCF-10A, MDA-MB-231, and A-MSC. Additionally, the expression levels of inflammatory cytokines (IL-1β, IL-6, IL-10, and TNF-α) were examined. Methylthiazolyldiphenyl-tetrazolium bromide assay demonstrated that free DOX showed the highest cytotoxicity against MCF-10A cells, followed by MCF-7 cells. Conversely, EXO-DOX indicated a greater effect on MCF-7 cells and had a lower IC50 compared to MDA-MB-231 cells. Free DOX significantly downregulated the expression of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), particularly in MCF-7 and MCF-10A cells, while concurrently upregulating IL-10 expression. EXO-DOX induced a more significant alteration in cytokine expression than the control and free DOX treatment groups. The co-culture system revealed a synergistic effect of free DOX on cancer cells while simultaneously mitigating the toxic effects of DOX on normal cells. This study suggests that EXO-DOX has promising potential as a targeted drug delivery system that could potentially improve therapeutic efficacy and minimize off-target toxicity.