Lung cancer, especially non-small cell lung cancer (NSCLC), is one of the most complex diseases, despite the existence of effective treatments such as chemotherapy and immunotherapy. Since cancer stem cells (CSCs) are responsible for chemo- and radio-resistance, metastasis, and cancer recurrence, finding new therapeutic targets for CSCs is critical. Dinactin is a natural secondary metabolite produced by microorganisms. Recently, dinactin has been revealed as a promising antitumor antibiotic via various mechanisms. However, the evidence relating to cell cycle progression regulation is constrained, and effects on cancer stemness have not been elucidated. Therefore, the aim of this study is to evaluate the new function of dinactin in anti-NSCLC proliferation, focusing on cell cycle progression and cancer stemness properties in Lu99 and A549 cells. Flow cytometry and immunoblotting analyses revealed that 0.1–1 µM of dinactin suppresses cell growth through induction of the G₀/G₁ phase associated with down-regulation of cyclins A, B, and D3, and cdk2 protein expression. The tumor-sphere forming capacity was used to assess the effect of dinactin on the cancer stemness potential in NSCLC cells. At a concentration of 1 nM, dinactin reduced both the number and size of the tumor-spheres. The quantitative RT-PCR analyses indicated that dinactin suppressed sphere formation by significantly reducing expression of CSC markers (i.e., ALDH1A1, Nanog, Oct4, and Sox2) in Lu99 cells. Consequently, dinactin could be a promising strategy for NSCLC therapy targeting CSCs.

Multidrug-resistant (MDR) bacteria, especially Escherichia coli, are a major contributor to healthcare-associated infections globally, posing significant treatment challenges. This study explores the efficacy of (−)-epigallocatechin gallate (EGCG), a natural constituent of green tea, in combination with ampicillin (AMP) to restore the effectiveness of AMP against 40 isolated MDR E. coli strains. Antimicrobial activity assays were conducted to determine the minimum inhibitory concentrations (MIC) of EGCG using the standard microdilution technique. Checkerboard assays were employed to assess the potential synergistic effects of EGCG combined with AMP. The pharmacodynamic effects of the combination were evaluated through time-kill assays. Outer membrane disruption was analyzed by measuring DNA and protein leakage and with assessments using N-phenyl-1-naphthylamine (NPN) and rhodamine 123 (Rh123) fluorescence dyes. Biofilm eradication studies involved biofilm formation assays and preformed biofilm biomass and viability assays. Scanning electron microscopy (SEM) was used to examine changes in cellular morphology. The results indicated that EGCG demonstrated activity against all isolates, with MICs ranging from 0.5 to 2 mg/mL, while AMP exhibited MIC values between 1.25 and 50 mg/mL. Importantly, the EGCG-AMP combination showed enhanced efficacy compared to either treatment alone, as indicated by a fractional inhibitory concentration index between 0.009 and 0.018. The most pronounced synergy was observed in 13 drug-resistant strains, where the MIC for EGCG dropped to 8 µg/mL (from 1 mg/mL alone) and that for AMP to 50 µg/mL (from 50 mg/mL alone), achieving a 125-fold and 1000-fold reduction, respectively. Time-kill assays revealed that the bactericidal effect of the EGCG-AMP combination occurred within 2 h. The mechanism of EGCG action includes the disruption of membrane permeability and biofilm eradication in a dose-dependent manner. SEM confirmed that the combination treatment consistently outperformed the individual treatments. This study underscores the potential of restoring AMP efficacy in combination with EGCG as a promising strategy for treating MDR E. coli infections.

Anticancer and Antimutagenic Properties of Pogonatherum paniceum on Colorectal Cancer Cells

Background: N-acetyl-p-aminophenol (APAP) overdose remains a leading cause of acute liver failure worldwide. The aril extracts derived from Gac (Momordica cochinchinensis) (MC) fruit contain diverse phytonutrients that exhibit a range of pharma-nutritional properties, including antioxidant and anti-hepatic damage properties. Despite the therapeutic potential, the molecular mechanisms underlying its hepatoprotective action, particularly in preventing drug-induced toxicity, remain unelucidated. Purpose: This study investigated the hepatoprotective potential of MC in a mouse model of APAP-induced acute liver injury and characterized its bioactive compounds. Methods: MC was prepared using aqueous extraction. Mice were pre-treated with either 500 or 1,000 mg/kg of MC for seven consecutive days prior to intraperitoneal administration of APAP 300 mg/kg to induce hepatotoxicity. Serum and liver samples were then collected to analyze biochemical changes, histopathological changes, oxidative stress markers, and/or mRNA expression. Standard chemical tests were used to identify bioactive compounds. Results: APAP administration caused a marked rise in liver enzymes, extensive histological necrosis, suppression of hepatocyte proliferation, infiltration of neutrophils and macrophages, and significant oxidative damage (p < 0.05). Studies revealed the constitution of MC as phenolic compounds, flavonoids, proanthocyanidins, and ascorbic acid, significantly attenuated the hepatotoxicity (p < 0.05). Mechanistically, MC ameliorated hepatic mRNA expression of CYP2E1, JNK, Ddit3, Bax, Casp3, NF-κB, and MCP-1, while increasing Bcl-2, IL-10, VEGF, Nrf2, SOD2, and GCLC mRNA (p < 0.05). Furthermore, MC prevented hyperglycemia by influencing the expression of AdipoR1, GLUT-2, and PEPCK (p < 0.05). Conclusion: MC exhibits hepatoprotective effects against APAP-induced liver injury through multifaceted modulation of toxic metabolism (CYP2E1), cell death and ER stress (JNK, Ddit3, Bax, Casp3, and Bcl-2), antioxidant response (Nrf2 pathway), inflammation (NF-κB pathway and IL-10), tissue repair (VEGF), and glucose metabolism (AdipoR1, GLUT-2, and PEPCK).

Background: Produced from Streptomyces, Dinactin is one of the most important macrolide antibiotics. Previous studies have found that dinactin exerts inhibiting the cancer stemness potential of lung cancer, making it a promising option for CRC therapy. In this study, we designed experiments to evaluate dinactin’s inhibitory effects against two human colorectal cancer cells (HCT116 and HT29) and elucidate the mechanism of its anti-cancer effects. Finally, we provided the first ever investigation of the mutagenic and anti-mutagenic potentials of dinactin. Methods: We studied the effects of dinactin as part of an anti-cancer approach using MTT test, clonogenic assay, wound healing assay, and transwell migration assay in HCT116 and HT29 cells. Mechanical studies were conducted using flow cytometry and Cell Death Detection ELISA. The mutagenic and anti-mutagenic properties were demonstrated using the in vitro cytokinesis-blocked micronucleus assay in V79 cells. Results: Dinactin was found to have dose-dependent inhibitory effects on the viability of HCT116 and HT29 cells with IC50 values of 1.6 ±0.4 and 35.0 ±14 nM, respectively. Furthermore, dinactin showed potent anti-proliferation, anti-migration, and anti-invasion activities and demonstrated similar activity to doxorubicin. Mechanically, treatment with dinactin resulted in increased DNA fragmentation and positive AnnexinV/FITC-PI staining, which demonstrated the induction of apoptosis by dinactin. From the results of the mutagenic assay, dinactin did not produce a significant increase in micronucleus frequency when compared with a negative control (p > 0.05). In order to verify dinactin’s anti-mutagenic properties, a combination of MMC (a known mutagen) and dinactin were applied to V79 cells. Treatment with dinactin resulted in a significant decrease in the number of micronucleated cells. Conclusion: Dinactin inhibits growth and alleviates the progression of CRC cells at nanomolar concentrations. This anti-CRC activity is mediated through the induction of apoptosis. Dinactin did not exhibit mutagenic effects and has the potential to reduce the mutagenic action of MMC.

Acute Lymphocytic Leukemia (ALL) is the most prevalent childhood cancer. Early diagnosis of ALL is essential, since timely and appropriate therapy significantly improves survival rates. ALL entails abnormal white blood cell (WBC) proliferation that weakens the immune system. Despite numerous studies on ALL identification using microscopic blood smear images, challenges remain due to the structural complexity of blood cells, image noise, intensity inhomogeneity, and cell overlap. Deep learning techniques, particularly CNNs, have shown high efficacy in medical image analysis. However, some studies employ GLCM features that lack an empirical basis for their selection. The study aims systematically evaluate nine distinct GLCM features to provide evidence-based reference for their use in ALL detection. Foundational CNN architecture was used as a classifier to establish a performance baseline with raw color images. Then we compared the baseline to models enhanced with each of the nine GLCM features using the public C-NMC 2019 dataset. The results revealed that GLCM Dissimilarity was the most effective feature, yielding an accuracy of 94.37% and an F1-score of 0.9437. The GLCM Dissimilarity significantly outperformed other GLCM features, followed by GLCM Entropy (93.84%), GLCM Mean (93.57%), GLCM Contrast (93.55%), GLCM Standard Deviation (93.13%), and GLCM ASM (90.98%). A model using only color images achieves an accuracy of 90.53%. It is important to note that although several GLCM features improved performance, among the nine features evaluated, those not listed among the top performers achieved lower accuracy than the color image baseline, highlighting the importance of careful feature selection. These results indicate that specific GLCM features, particularly GLCM Dissimilarity, can substantially enhance CNN-based ALL classification, underscoring its potential as a robust image descriptor for diagnostic applications.