Hana Hammad

Background/Objectives: Doxorubicin (DOX) is an effective chemotherapeutic agent, but its clinical use is limited by dose-dependent cardiotoxicity. Emerging evidence suggests that epigenetic dysregulation, particularly altered DNA methylation, contributes to DOX-induced cardiac injury. Metformin has been reported to exert cardiometabolic and epigenetic regulatory effects. This study investigated genome-wide DNA methylation changes induced by chronic metformin exposure and their effects on doxorubicin sensitivity in H9c2 cardiomyoblast cells. Methods: Genome-wide DNA methylation changes induced by chronic metformin exposure were investigated in H9c2 cardiomyoblast cells using whole-genome bisulfite sequencing (WGBS). Cells were treated with metformin (0.7–2.8 mM) for four months prior to DOX exposure. Cellular sensitivity to DOX was evaluated using MTT-based dose–response analysis and IC₅₀ estimation. Results: DOX reduced cell viability (IC₅₀ = 0.164 µM). Chronic metformin pre-treatment produced a dose-dependent rightward shift in DOX dose–response curves, increasing IC₅₀ values to 0.21, 0.289, and 0.51 µM at 0.7, 1.4, and 2.8 mM metformin, respectively. WGBS revealed distinct separation between treatment groups in principal component analysis. Significant methylation changes (adjusted p-value < 0.05) were identified in genes related to oxidative stress, mitochondrial function, apoptosis, and chromatin regulation. Conclusions: Chronic metformin exposure induces dose-dependent genome-wide DNA methylation remodeling in cardiac cells and is associated with altered cellular sensitivity to doxorubicin. These findings suggest that metabolic modulation by metformin may influence epigenetic regulation and cellular stress responses relevant to chemotherapy-induced cardiotoxicity.

Background: Doxorubicin causes dose-dependent cardiotoxicity linked to epigenetic dysregulation, especially DNA methylation. Metformin shows cardioprotective effects through metabolic and epigenetic mechanisms. This study examined the role of metformin in counteracting doxorubicin-induced DNA methylation changes. Methods: H9c2 cardiomyoblasts were treated with doxorubicin, with or without metformin (0.7–2.8 mM). Cell viability and IC₅₀ were determined by MTT assay. Genome-wide DNA methylation was analysed by whole-genome bisulfite sequencing, followed by PCA and differential methylation analysis with FDR correction. Results: Doxorubicin reduced cell viability (IC₅₀ = 0.164 μM). Pre-treatment with metformin increased IC₅₀ to 0.21, 0.289, and 0.51 μM (0.7, 1.4, 2.8 mM). PCA showed distinct separation among groups. Numerous differentially methylated regions (FDR < 0.05) were associated with genes involved in oxidative stress, mitochondria, apoptosis, and chromatin regulation. Conclusions: Metformin induced distinct, dosedependent genome-wide DNA methylation changes in cardiac cells, indicating a direct epigenetic effect. These findings suggest that metformin influences cardiac epigenetic regulation and may contribute to cardioprotection by modulating DNA methylation.

Background: P2 receptors have been implicated in the release of neurotransmitter and pro-inflammatory cytokines due to their response to neuroexcitatory substances in the microglia. The P2X4, P2X7 and P2Y12 receptors are involved in the development of pain behavior induced by peripheral nerve injury. However, it is not known if blocking P2X4, P2X7 and P2Y12 receptors is associated with the expression and the release of interleukin-1B (IL-1β), interleukin-6 (IL-6), or tumor necrosis factor-α (TNF-α) in cultured neonatal spinal cord microglia. Objective: For this reason, we examined the effects of P2X4, P2X7 and P2Y12 antagonists on the expression and the release of IL-1β, IL-6, and TNF-α in ATP-stimulated microglia. Methods: In this study, we observed the effect of A-740003, PSB-12062 and MRS 2395 (P2X4, P2X7 and P2Y12 receptors antagonist, respectively), on the expression and release of IL-1β, IL-6 and TNF-α by using real-time fluorescence quantitative polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Results: ATP induced the increased expression of IL-1β, IL-6 and TNF-α at the level of messenger RNA (mRNA). ATP-evoked increase in IL-1β, IL-6 and TNF-α mRNA expression was inhibited by the P2X4 receptor antagonist A-740003 or P2X7 receptor antagonist PSB-12062, respectively. Similarly, ATP-evoked release of IL-1β, IL-6 and TNF-α was inhibited by A-740003 and PSB-12062. Furthermore, ATP-evoked increased expression of Iba-1, IL-1β, IL-6 and TNF-α mRNA, and release of IL-1β, IL-6 and TNF-α were nearly all blocked after co-administration of A-740003 plus PSB-12062. Finally, ATP-evoked increased gene expression and release of IL-1β, IL-6 and TNF-α were also inhibited by MRS 2395 (P2Y12 antagonist). Conclusion: These observations suggest a new clue for therapeutic strategies to treat the neuro-inflammation. Pharmacy Practice is a free full-text journal with a scope on the practice of pharmacy. Pharmacy Practice covers pharmaceutical care, social pharmacy, pharmacy education, process and outcome research, health promotion and education, health informatics, pharmacoepidemiology

Objective: Achillea fragrantissima L. (Asteraceae) is a traditionally used medicinal herb in the rural communities of Jordan. Methods: The present study evaluated the efficacy of the ethanol extract of this species on angiogenesis in both, ex vivo using rat aortic ring assay and in vivo using rat excision wound model. Results: In concentrations of 50 and 100 µg/ml, the ethanol extract showed angiogenic stimulatory effect and significantly increased length of capillary protrusions around aorta rings of about 60% in comparison to those of untreated aorta rings. In MCF-7 cells, the ethanol extract of A. fragrantissima stimulates the production of VEGF in a dose-dependent manner. 1% and 5% of ethanol extract of A. fragrantissima containing vaseline based ointment was applied on rat excision wounds for six days and was found to be effective in wound healing and maturation of the scar. Both preparations resulted in better wound healing when compared to the untreated control group and vaseline-treated group. This effect was comparable to that induced by MEBO, the positive control. Conclusion: The results indicate that A. fragrantissima has a pro-angiogenic effect, which may act through the VEGF signaling pathway.

The development of plant‐based nano‐materials is considered an eco‐friendly technology because it does not involve hazardous chemicals. In this study, bimetallic ZnFe₂O₄ and CrFe₂O₄ nanoparticles were synthesized using an aqueous extract of Boswellia carteri resin. Synthesized ZnFe₂O₄ and CrFe₂O₄ nanoparticles were characterized by UV‐Vis spectroscopy, FTIR, XRD, and HR–TEM. The anti‐inflammatory activity was investigated in LPS‐stimulated RAW 264.7 macrophages, whereas antioxidant activity was examined using a Hydrogen Peroxide Scavenging Activity Assay, Nitric Oxide Scavenging Activity Assay, and ABTS Radical Scavenging Assay. ZnFe₂O₄ and CrFe₂O₄ nanoparticles demonstrated a moderate scavenger of H₂O₂ with IC₅₀ values; 87.528 ± 8 μg/ml and 146.4468 ± 12 μg/ml, respectively. While they exhibited a strong scavenger of NO with IC₅₀ values; 4.01 ± 0.7 μg/ml and 4.01 ± 0.7μg/ml, respectively. Interestingly, ZnFe₂O₄ and CrFe₂O₄ nanoparticles revealed an excellent anti‐inflammatory activity by dose‐dependently suppressing mRNA expressions of IL‐1b, IL‐6, and TNF‐α. Also, ZnFe₂O₄ and CrFe₂O₄ nanoparticles suppress the protein expression of TNF‐α. Together, our results proved that phyto‐mediated ZnFe₂O₄ and CrFe₂O₄ nanoparticles using Boswellia carteri resin have great potential in biomedical applications such as anti‐inflammatory and antioxidant. Practical applications Our phyto‐synthesized chromium iron oxide bimetallic nanoparticles (NPs) have shown a novel and potent anti‐inflammatory activity, with remarkable biosafety toward tested macrophages. Zinc iron oxide bimetallic NPs exhibited anti‐inflammatory effect with a lesser extent compared to the former, with moderate cytotoxicity against tested macrophages. Both zinc and chromium iron oxide NPs exhibited an equivalent antioxidant activity. Our resin‐capped chromium iron oxide NPs are suggested to be a competing nonsteroidal anti‐inflammatory agent; it is further recommended to establish advanced animal studies to confirm their biosafety, stability, and anti‐inflammatory activity accompanied with the antioxidant activity.

Objective: This study aims to identify the changes in expression of hypoxia-inducible genes in seven different cancer cell lines that vary in their oxygen levels in an attempt to identify hypoxia biomarkers that can be targeted in therapy. Profiling of hypoxia inducible-gene expression of these different cancer cell lines can be used as a baseline data for further studies. Methods: Human cancer cell lines obtained from the American Type Culture Collection were used; MCF7 breast cancer cells, PANC1 pancreatic cancer cells, PC-3 prostate cancer cells, SH-SY5Y neuroblastoma brain cancer cells, A549 lung cancer cells, and HEPG2 hepatocellular carcinoma. In addition, we used MCF10A non-tumorigenic human breast epithelial cell line as a normal cell line. The differences in gene expression were examined using real-time PCR array (PAHS-032Z, Human Hypoxia Signaling Pathway PCR Array) and analyzed using the ΔΔCt method. Results: Almost all hypoxia-inducible genes showed a PO2-dependent up- and down-regulated expression. Noticeable gene expression differences were identified. The most important changes occurred in the HIF1α and NF-KB signaling pathways targeted genes and in central carbon metabolism pathway genes such as HKs, PFKL, and solute transporters. Conclusion: This study identified possible hypoxia biomarkers genes such as NF-KB, HIF1α, HK, PFKL, and PIM1 that were expressed in all hypoxic cells. Pleotropic pathways that play a role in inducing hypoxia directly such as HIF1 α and NF-kB pathways were upregulated. In addition, genes expressed only in the severe hypoxic liver and pancreatic cells indicate that severe and intermediate hypoxic cancer cells vary in their gene expression. Gene expression differences between cancer and normal cells showed the shift in gene expression profile to survive and proliferate under hypoxia.

Four new complexes derived from adamantly containing hydrazone (APH) ligand with Cu(II) (1), Co(II) (2), Ni(II) (3) and Zn(II) (4), have been synthesized and characterized using different physicochemical methods. The structure of the ligand APH and its copper complex 1 have been established by single-crystal X-ray diffraction direct methods, which reveal that complex 1 has distorted square-pyramidal geometry. Complexes 1–4 are screened against seven human cancer cell lines namely, breast cancer cell lines (MCF7, T47D, MDA-MB-231), prostate cancer cell lines (PC3, DU145) and the colorectal cancer cell line Coco-2, for their antiproliferative activities. Complex 1 has shown a promising anticancer activity compared to the other ones. The structural and spectroscopic analysis of APH and its complexes are confirmed by DFT calculations.

Background: Cytochrome P450 2A6 enzyme (CYP2A6), an essential hepatic enzyme involved in the metabolism of drugs, is responsible for a major metabolic pathway of nicotine. Variation in the activity of polymorphic CYP2A6 alleles has been implicated in inter-individual differences in nicotine metabolism. Aims: The objective of the current study was to assess the association between the smoking status and the cytochrome P450 2A6 enzyme (CYP2A6) genotype in Jordanians. Methods: In the current study, 218 (117 Male and 101 female) healthy unrelated Jordanian volunteers were recruited. CYP2A6*1B, CYP2A6*4 and CYP2A6*9 were determined and correlated with subject smoking status. Results: *1A/*1A was the most common genetic polymorphism in the overall study population, with no significant frequency differences between smokers and non-smokers. When the population was divided according to gender, only male smokers showed a significant correlation between genotype and smoking status. Considering the CYP2A6*9 genotype, the results showed differences in distribution between smokers and non-smokers, but only women showed a significant association between CYP2A6*9 allele genotype and smoking status. Conclusion: The results of this study show that there is a significant association between CYP2A6*9 genotype and smoking status. They also show that CYP2A6 genotype is significantly influenced by gender.