Faculty of Biotechnology Research Paper

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FokI polymorphism of the vitamin D receptor gene: Linking COVID-19 risk to genetic susceptibility in children
(Academic Press, 2025-05-13) Amal Ahmed Mohamed; Abdullah Taher Alanazi; Hoda H. Ahmed; Samar Elfiky; Muhammad T Abdel Ghafar; Ingy Maher; Sherin A. Taha; Mohammed Zakaria Ali Abu Rahma; Waleed Elagawy; Dina A. Mohareb; Abeer M. Rawy; Heba M. Abostate; Amira AlSayed Youssef; Dalia Saeed Elsayed; Rasha M. Abdel-Hamid
Background Vitamin D receptor (VDR), influenced by gene polymorphisms like FokI, may affect susceptibility to infections, including coronavirus disease 2019 (COVID-19). Since studies in children are limited, we aimed to analyze the correlation between the VDR FokI variant and both the incidence and severity of COVID-19 in Egyptian children. Methods Seventy-seven COVID-19-positive and 107 COVID-19-negative pediatric patients were included. Participants' serum 25(OH)D levels, inflammatory biomarkers, and demographics were evaluated. Real-time polymerase chain reaction (PCR) was used for genotyping the VDR FokI (rs2228570) polymorphism. Results Absolute lymphocyte count (ALC) was significantly lower in COVID-19 patients than in controls, while interleukin-6 (IL-6), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), procalcitonin, and D-dimer were significantly higher (all p < 0.001). Vitamin D insufficiency was significantly more common in COVID-19 cases (18.2 % versus 3.7 %, p = 0.002). Male sex, increased tumor necrosis factor-alpha (TNF-α), and CRP were significantly associated with severe COVID-19 (p = 0.032, 0.029, < 0.001, respectively). The FokI TT genotype in codominant and recessive models and the T allele in the multiplicative model were significantly correlated with 2.4, 3.0, and 1.8 folds increased COVID-19 risk (p = 0.043, < 0.001, and 0.004, respectively). However, VDR FokI variants did not significantly associate with severe COVID-19. Conclusion The T allele and TT genotype of the FokI variant in the VDR gene increase susceptibility to COVID-19 but not its severity in Egyptian children. Additional research is required to validate the potential role of vitamin D and its receptor polymorphism in COVID-19.
Caffeine-boosted silver nanoparticles target breast cancer cells by triggering oxidative stress, inflammation, and apoptotic pathways
(Elsevier B.V., 2025-05-14) Naief Dahran; Mohamed S. Othman; Farah Mumtaz; Ghada M. Aleid; Mohamed E. Ghoniem; Mai A. Samak; Mohamed T. Elabbasy; Ayman A. Saleh; Sofian T. Obeidat; Ola A. Habotta; Ahmed E. Abdel Moneim
Breast cancer (BC) constitutes a major global health concern and is the second foremost cause of cancer-related mortality among women worldwide. This research investigated the anticancer effectiveness of caffeine-conjugated silver nanoparticles (Caf-AgNPs) against MDA-MB-231 breast cancer cells, utilizing fluorouracil (5-FU) as a reference antitumor drug. The study illustrated that the strategic conjugation of caffeine with AgNPs substantially improved the therapeutic efficacy against breast cancer cell lines and simultaneously attenuated cytotoxicity in normal mouse liver (NBL) cells. Caf-AgNPs significantly increased ROS, malondialdehyde, COX-2, IL-1β, and TNF-α level in BC cells, which was accompanied by a decrease in glutathione levels. The increased levels of cytosolic cytochrome c, caspase-3, and Bax proteins, as well as a significant decrease in Bcl-2 expression and Bcl-2/Bax ratio, were indicative of the significant pro-apoptotic effects of Caf-AgNPs in MDA-MB-231 cells. Cancer cells subjected to Caf-AgNPs demonstrated elevated lactate dehydrogenase (LDH) membrane leakage, signifying cellular membrane disruption. Cell cycle analysis revealed a substantial proportion of early and late stage apoptosis in cancer cells exposed to Caf-AgNPs, accompanied by a notable downregulation of cyclin D1 and cyclin-dependent kinase 2 (CDK2) mRNA expression. Caf-AgNPs utilize several mechanisms for cellular destruction, including cell cycle arrest, oxidative stress induction, modulation of the inflammatory response, and mitochondrial apoptosis. Caf-AgNPs offer a promising and complex strategy for breast cancer intervention. © 2025 American Pharmacists Association
Synthesis of rice husk-loaded chitosan nanoparticles for treatment of amikacin-induced kidney injury in male rats
(Elsevier B.V, 2025-05-10) Alyaa Farid; Aisha Hossam; Malak Waheed; Gehan Safwat
The main causes of amikacin-induced kidney damage are inflammation and oxidative stress. Although rice husks are full of bioactive components that have therapeutic uses, their instability and low bioavailability limit their use. Chitosan nanoparticles improve the stability and bioactivity of phytochemicals by offering an effective drug delivery method. The study aimed to prepare and characterize rice husk-loaded chitosan NPs and evaluate their therapeutic effects against amikacin-induced nephrotoxicity. Ionic gelation was used to create NPs, which were then examined using transmission electron microscopy, zeta potential, and dynamic light scattering. In vitro tests were performed to evaluate their anti-inflammatory and antioxidant properties. Male rats were divided into: negative control, positive control, amikacin-induced kidney injured rats treated with rice husks extract, chitosan NPs or rice husk-loaded chitosan NPs. Gas chromatography verified that rice husks phytochemicals were successfully loaded on chitosan NPs. Rice husk-loaded chitosan NPs had a consistent zeta potential (~58.6 mV) and a nanoscale size (~76.7 nm). When compared to rice husk extract or chitosan NPs, rice husk-loaded chitosan NPs showed better anti-inflammatory and antioxidant properties. In vivo, rice husk-loaded chitosan NPs significantly restored kidney function biomarkers, lowered oxidative stress and inflammation. In conclusion, rice husk-loaded chitosan NPs is a nephroprotective treatment.
A Guide to Basic RNA Sequencing Data Processing and Transcriptomic Analysis
(Bio-protocol LLC, 2025-05-05) Rowayna Shouib; Gary Eitzen; Rineke Steenbergen
RNA sequencing (RNA-Seq) has transformed transcriptomic research, enabling researchers to perform largescale inspection of mRNA levels in living cells. With the growing applicability of this technique to many scientific investigations, the analysis of next-generation sequencing (NGS) data becomes an important yet challenging task, especially for researchers without a bioinformatics background. This protocol offers a beginner-friendly step-by-step guide to analyze NGS data (starting from raw .fastq files), providing the required codes with an explanation of the different steps and software used. We outline a computational workflow that includes quality control, trimming of reads, read alignment to the genome, and gene quantification, ultimately enabling researchers to identify differentially expressed genes and gain insights on mRNA levels. Multiple approaches to visualize this data using statistical and graphical tools in R are also described, allowing the generation of heatmaps and volcano plots to represent genes and gene sets of interest.
Metabolomic analysis reveals key changes in amino acid metabolism in colorectal cancer patients
(Springer, 2025-05-02) Asmaa Ramzy; Taghreed Khaled Abdelmoneim; Menna Arafat; Maha Mokhtar; Ashraf Bakkar; Amany Mokhtar; Wagida Anwar; Sameh Magdeldin; Shymaa Enany
The number of colorectal cancer (CRC) patients is steadily growing worldwide, particularly in developing nations. Nonetheless, recent advances in early detection studies and therapy alternatives have reduced CRC mortality in afuent countries, despite rising incidence. Gut microbiota and their metabolites may contribute to tumor growth and reduced therapeutic efcacy. This preliminary study sought to uncover metabolic fngerprints in colorectal cancer patients. It also emphasizes the correlation between the gut microbiome, microbial metabolism, and altered metabolites in CRC. In this study, stool samples from 20 CRC patients and matched healthy controls were enrolled. Untargeted metabolomics approach based on an ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-MS/MS) were applied. Statistical approaches, pathway enrichment analysis, and network analysis were employed to unleash CRC perturbed metabolic pathways and putative biomarkers. The study identifed a distinct manually curated metabolite profle that is substantially linked to CRC. The steroidogenesis, aspartate, tryptophan (Trp), and urea cycle were the most signifcant pathways that concurrently contributed to CRC.Prominently, among other pathways, Trp metabolism was identifed as a critical pathway, indicating a possible connection between the development of CRC and gut microbiota. In a nutshell the notable resulted metabolites reveal auspicious biomarkers for the initial diagnosis as well as surveilling of CRC progression. This preliminary study highlights the potential involvement that gut bacteria may contribute in CRC patients. Further investigation into the composition of the gut microbiome associated with this metabolic profle may lead to the identifcation of novel biomarkers for early detection and possible targets for treatment.
Freshwater Bivalve Coelatura aegyptiaca as a Sensitive Bioindicator for Zinc Oxide/ Chitosan Nanocomposites Toxicity
(Bentham Science Publishers, 2025-04-11) Mennatallah H. Abdelaziz; Muhammed S. ElRakabawi; Ahmed Mohamed Soliman; Ayman Saber Mohamed
Background: Freshwater bivalves are considered effective biomarkers for pollution in aquatic ecosystems. Despite advances in the use of zinc oxide nanoproducts in several sectors, such as food, industry, and medicine, paying attention to their environmental impacts is crucial. The objective of the study was to investigate the mechanisms of zinc oxide-chitosan nanocomposites (ZnO-CS NCs) ecotoxic in freshwater environments using freshwater bivalves Coelatora aegyptica as a sensitive indicator. Methods: After preparing and characterizing ZnO NPs and ZnO-CS NCs with transmission electron microscopy, ultraviolet spectroscopy, and X-ray diffraction, we exposed the bivalve to three different doses of ZnO NPs and ZnO-CS NCs (12.5, 25, and 50 mg/L) for 7 days. Results: ZnO-CS NCs concentrations significantly increased malondialdehyde and nitric oxide levels, whereas glutathione and catalase levels decreased in investigated organs. Furthermore, histological changes were detected in the tissues of the gills and mantle. Discussion: The bivalve organs had varying quantities of MDA, NO, GSH, and CAT. This could be related to the accumulation pattern of heavy metals in each organ, their close interaction with water, or the removal rates. Conclusion: We concluded from our findings that the toxicity of ZnO-CS NCs on freshwater bivalves causes histological alterations and an oxidative stress response. Moreover, Coelatura aegyptiaca was proposed as a highly sensitive bioindicator to monitor water contamination induced by diverse nanoparticles because it can accumulate and concentrate most pollutants, even at low concentrations. As a result, we recommend conducting additional studies with fresh bivalves to evaluate the aquatic ecosystem well and reduce water contamination at both local and worldwide levels.
Gamma radiation-assisted synthesis of tea tree oil-based chitosan films for active packaging applications
(John Wiley and Sons Ltd, 2025-04-13) Asmaa Sayed; Manar E. Abdel-Raouf; Mikhail Magdy; Ghada A. Mahmoud
Sustainable packaging materials with inherent antimicrobial and antioxidant properties are increasingly demanded for food preservation. In this study, chitosan/polyacrylamide/tea tree oil (CS/PAAm/TTO) films were developed via gamma irradiation, varying both the irradiation dose (10–50 kGy) and the TTO concentration (25–100 μL) to attain an optimized formulation with superior performance. In this regard, the optimization was accomplished via atomic force microscopy (AFM) through assessing the surface morphologies of the films prepared at different irradiation doses, which identified 30 kGy as the optimal irradiation dose for TTO incorporation. On the other hand, comprehensive characterizations using Fourier transfer infrared spectroscopy, x-ray diffraction, thermogravimetric analysis, and contact angle measurements revealed that increasing the irradiation dose and TTO content enhanced hydrophobicity and improved mechanical properties. More specifically, the contact angle increased significantly at higher TTO levels, indicating an increased surface hydrophobicity, while tensile tests demonstrated improved flexibility. In addition, antimicrobial assays confirmed the efficacy of the films against a range of pathogens, and the 1,1-diphenyl-2-picryl hydrazyl assay indicated notable antioxidant activity, with an IC50 value of 111.45 μg/mL for the optimized formulation. These findings underscore the potential of the developed biofilms as advanced, multifunctional packaging materials, offering a promising environmentally friendly alternative to conventional packaging for food preservation.
Synthesis, structural characterization, in vitro biological evaluation and in silico molecular docking of some new 1,2,3-Benzotriazole-based Schiff base derivatives
(Elsevier B.V, 2025-04-09) Shawkat Hayat; Mohamed S. Othman; Hayat Ullah; Amina Qureshi; Fazal Rahim; Sadiqa Begum; Mohsan Nawaz; Abdul Wadood; Amal H. Al-Bagawi; Ahmed M. Aref; Mohamed A. Fareid; Rashid Iqbal
1,2,3-benzotriazole-based Schiff base derivatives were synthesized, characterized through different techniques such as 1 HNMR, 13CNMR, and HREI-MS, and screened against the alpha-glucosidase enzyme. All the synthesized analogues showed good inhibitory potentials with IC50 values ranging from 2.20 ± 0.30 μM to 24.40 ± 0.10 μM as compared to the standard drug acarbose (IC50 = 10.30 ± 0.20 μM). The most potent analogues among the series are 3 (IC50 = 4.30 ± 0.20 μM), 8 (IC50 = 9.40 ± 0.10), 9 (IC50 = 6.20 ± 0.40), 10 (IC50 = 6.60 ± 0.20), 11 (IC50 = 6.10 ± 0.30), 12 (IC50 = 4.70 ± 0.50 μM), 13 (IC50 = 2.20 ± 0.30 μM), 14 (IC50 = 4.30 ± 0.20 μM), and 16 (IC50 = 7.40 ± 0.20), which were found manyfold more active than the standard drug acarbose. A structureactivity relationship study was established that significantly depends on the position, nature, number, and electron-donating/withdrawing effect of the substituent(s) attached to the phenyl ring. We conducted molecular docking studies to investigate the binding interaction of the most potent analogues with the active site of an enzyme.
Modulating Effect of Green Tea and Vitamin C in the Management of “Alzheimer's disease: Oxidative Stress, Cell-Based Anti-Inflammatory and Genotoxic Profiles
(NIDOC (Nat.Inform.Document.Centre), 2025-04-16) Salwa M. El-Hallouty; khaled Mahmoud; Mahmoud Kh. Hanafy; Eman Ayman Abdellatef; Elham M.Youssef
Alzheimer's disease is a progressive brain disorder. Green tea (GT) and vitamin C are known for their antioxidant and anti-inflammatory properties. This study aims to investigate the potential oxidative stress, cell-based anti-inflammatory and genotoxic Profiles for synergistic effect of green tea and vitamin c against to Alzheimer's disease. In vitro assessments included their several antioxidant parameters and ability to inhibit inflammatory markers such as nitric oxide (NO), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and matrix metallopeptidase 1(MPP1) in lipopolysaccharide-activated macrophages. In vivo approach evaluated genotoxicity and their potential neurotherapy against Alzheimer's disease using a rat model. Behavioral, biochemical, and histological evaluations were conducted. Green tea methanolic extract (GTME) and vitamin C showed enhanced antioxidant and anti-inflammatory modulator activities. The neurotoxicity rat’s model exhibited cognitive decline and brain injury in contrast to the control group. While treated rats with GTME and vitamin C exhibited improved cognitive function, behavior and reduced AChE levels the combination exerted a significantly more potent effect. Histological analysis confirmed improved brain tissue structure in rats receiving the combined treatment. Crucially, neither compound exhibited genotoxic properties. The combination of GTME and vitamin C demonstrated a promising therapeutic potential for neurodegenerative diseases due to their synergistic effects.
Potent cytotoxicity and induction of ROS‑mediated genomic instability, mitochondrial dysfunction, and apoptosis by Y2O3 NPs in Hep‑G2 hepatic cancer cells
(Springer Science and Business Media Deutschland GmbH, 2025-04-10) Hanan R. H. Mohamed; Rawan Essam; Basma A. Mohamed; George M. Hakeem; Shahd H. Elnawasani; Maria Nagy; Gehan Safwat; Ayman Diab
Hepatic cancer, one of the most prevalent and lethal cancers globally, remains a signifcant health challenge, with limited treatment options underscoring the urgent need for novel, more efective therapies. Yttrium oxide nanoparticles (Y2O3 NPs) have attracted attention in nanomedicine due to their promising properties, including enhanced drug delivery, imaging capabilities, and therapeutic efects. However, the specifc impact of Y2O3 NPs on hepatic cancer is largely unexplored. Therefore, this study was conducted to assess the cytotoxic efects of Y2O3 NPs on cell viability, reactive oxygen species (ROS) generation, genomic stability, mitochondrial integrity, and apoptosis induction in Hep-G2 hepatic cancer cells. The results from the SRB cytotoxicity assay demonstrated a strong concentration-dependent decrease in Hep-G2 cell viability, with a notably low half-maximal inhibitory concentration (IC50) value of 13.15 µg/ml. Exposure to the IC50 concentration of Y2O3 NPs led to increased ROS generation, DNA damage induction, and loss of mitochondrial membrane potential. Furthermore, the expression of pro-apoptotic p53 and mitochondrial ND3 genes was signifcantly upregulated, while the anti-apoptotic Bcl-2 gene was markedly downregulated, triggering apoptosis in Hep-G2 cells after 72 h of exposure to Y2O3 NPs. Collectively, these fndings highlight the therapeutic potential of Y2O3 NPs in hepatic cancer, emphasizing the need for further research to fully explore their efcacy as a treatment option for liver cancer
Towards propagation of epidermal cells for wound repair: glass, as cell culture substrate, enhances proliferation and migration of human keratinocytes
(Frontiers Media SA, 2025-03-20) Hady Shahin; Ingrid Steinvall; Folke Sjöberg; Moustafa Elmasry; Ahmed El-Serafi
Introduction: Human keratinocytes require relatively long propagation time which impedes their availability as autologous cell transplantation within a clinically reasonable timeframe. There is an unmet need for efficient xenofree cell expansion approaches to propagate human keratinocytes as regenerative therapy. Methods: Primary human keratinocytes and HaCaT cells were cultured on glass, plastic, and animal-derived collagen I matrix for 10 days. Proliferation, migration, DNA methylation, as well as gene and protein expression were assessed to characterize the effect of the tested culture substrates on keratinocytes at the molecular and functional levels. Results: Keratinocytes cultured on glass exhibited faster proliferation, global DNA demethylation and upregulation of epidermal differentiation markers. Scratch wound assay revealed that keratinocytes cultured on glass demonstrated enhanced cell migration compared to those on plastic or collagen I. Multiplex immunoassays identified temporal and substrate-dependent variations in a panel of keratinocyte-specific secreted factors, encompassing immunomodulatory cytokines, growth factors, and angiogenic factors. Discussion: Glass, as a culture substrate, promotes epidermal differentiation and enhances keratinocyte migration. The latter is a critical factor in reepithelialization and wound healing. Functional properties suggest that glass may optimize the inflammatory response and promote efficient wound repair, making it a promising candidate for the short-term expansion of keratinocytes for transplantation purposes. Further in-vivo validation is required to definitively establish the efficacy of keratinocytes cultured on glass for clinical applications.
EXT1 as an Independent Prognostic Biomarker in Breast Cancer: Its Correlation with Immune Infiltration and Clinicopathological Parameters
(Multidisciplinary Digital Publishing Institute (MDPI), 2025-03-13) Amira Hossny; Hatem A. F. M. Hassan; Sherif Ashraf Fahmy; Hazem Abdelazim; Mahmoud Mohamed Kamel; Ahmed H. Osman; Sherif Abdelaziz Ibrahim
Exostosin 1 (EXT1) encodes a type II transmembrane glycosyltransferase residing in the endoplasmic reticulum and plays an essential role in the elongation of heparan sulfate chain biosynthesis. Additionally, EXT1 may act as an oncogene that could promote cell proliferation as well as cancer cell metastasis. Herein, we investigated EXT1’s expression pattern and prognostic value in breast cancer, along with its immunological implications. Immunohistochemical staining of EXT1 was assessed in 85 breast cancer patients. Patients were categorized into molecular subtypes, namely luminal A, luminal B, and human epidermal growth factor receptor 2 (HER2), along with triple-negative breast cancer (TNBC). Correlations of EXT1 immunostaining with clinicopathological parameters were evaluated. Furthermore, the correlations of EXT1 expression with tumor immune infiltration and immune cell surface markers were assessed using TIMER. Moreover, survival analysis was conducted to reveal EXT1’s prognostic value. EXT1 expression was markedly associated with the status of the estrogen receptor (ER), molecular subtypes, and recurrence status. In addition, high levels of EXT1 expression were associated with worse overall survival (OS) and relapse-free survival (RFS). Analysis of immune infiltration indicated that EXT1 expression was positively correlated with dendritic cells (DCs), macrophages, neutrophils, CD4+ T cells, and CD8+ T cells, although it showed a negative correlation with the tumor purity. Overall, this study suggests that the elevated EXT1 expression, particularly in TNBC, has a positive correlation with poor prognosis and with immune-infiltrated cells in breast cancer. Therefore, it may emerge as an independent prognostic biomarker, immunological marker, and potential future therapeutic target for the most aggressive TNBC subtype.
Extraction of grape seeds by different solvents affects the activities of the resultant extract
(Springer Open, 2025-03-19) Kareem Tarek; Alyaa Farid; Gehan Safwat
Phenolic compounds are concentrated in grape seeds; 60–70% of the extractable grape phenols are found in the seeds. The focus of this research was to isolate the phytochemicals from grape seed and to determine their ability to prevent haemolysis, their antioxidant and microbiological activities. By using the extraction procedure, three solvents were used (distilled water, ethanol and methanol). A high-performance liquid chromatographic (HPLC) test was performed to analyse the phenolic compounds and flavonoids content that were used to determine the efficiency of the various solvents used in the extraction process. All the variables under study, namely yield percentage, phenolic component concentration, and flavonoid content got significantly affected by the choice of the solvent used. The flavonoid content of the extracts was in the order methanolic extract>ethanolic extract>water extract. The methanolic extract of the grape seeds exhibited the most powerful antioxidant and hemolysis inhibitory effects among the three extracts, followed by the ethanolic and water extracts. The antibacterial activity of methanolic extract was found to be higher as compared to the ethanolic extract against Staphylococcus aureus. The antibacterial activity of the ethanolic and methanolic extracts against Salmonella enteritidis, Bacillus subtilis, Aspergillus niger and Escherichia coli were found to be equivalent. In conclusion, grape seeds contained several bioactive compounds that exerted an antioxidant, hemolysis inhibition and anti-microbial activities. These activities depends on the concentration of phenolic compounds and flavonoids in the grape seed extracts. Methanol was the superior solvent in the extraction process followed by ethanol.
Resilient STR loci under gamma radiation: A preliminary study on DNA stability in buccal swabs
(Elsevier Ltd, 2025-03-06) Amani Kraishan; Ahmad M. Refaat; Jenan A. Almhaini; Alaa Saeed Bazuhair; Saeed M. Al-Qahtani; Marwan A. Althomali; Hatem Al-Ameryeen; Ali H. Alomari; Abdul-Wali Ajlouni
This study explores how gamma radiation affects DNA concentration and the stability of autosomal short tandem repeat (STR) markers in buccal swab samples. A total of 160 swabs from 20 volunteers were exposed to therapeutic radiation doses ranging from 1 to 80 Gy. The results revealed a clear dose-dependent decrease in DNA concentration, indicating significant DNA damage as radiation increased. Despite this, the autosomal STR markers remained stable across all doses, demonstrating their robustness against radiation-induced damage. These findings are crucial for clinical and forensic applications, confirming that STR profiling remains reliable for cancer patients undergoing radiation therapy and in forensic cases involving irradiated DNA. The study emphasizes the value of STR markers in genetic analysis under challenging conditions and highlights the need for further research on their long-term stability in diverse biological contexts.
Erbium oxide nanoparticles induce potent cell death, genomic instability and ROS‑mitochondrial dysfunction‑mediated apoptosis in U937 lymphoma cells
(Springer Verlag, 2025-03-22) Hanan R. H. Mohamed; Yusuf Ahmed Elberry; Hagar Magdy; Maryam Ismail; Maivel Michael; Nourhan Eltayeb; Gehan Safwat
Erbium oxide nanoparticles (Er2O3-NPs) have attracted signifcant attention for their unique physicochemical properties, including high surface area, biocompatibility, and stability. However, the impact of Er2O3-NPs on lymphoma cells (LCs) has not been explored, making this an innovative avenue for exploration. Therefore, the current study aimed to explore the infuence of Er2O3-NPs on cell viability, genomic and mitochondrial DNA integrity, reactive oxygen species (ROS) generation and apoptosis induction in human U937 LCs. Er2O3-NPs were characterized using X-ray difraction (XRD) and transmission electron microscopy (TEM). The efect of Er2O3-NPs on cell viability and genomic DNA integrity was estimated after 48 h using the WST-1 cytotoxicity and alkaline Comet assays, respectively. The generation level of reactive oxygen species (ROS) and mitochondrial membrane potential were also analyzed. Flow Cytometry was used to assess apoptosis induction and quantitative RT-PCR was conducted to measure the apoptotic (p53), anti-apoptotic (Bcl2), and mitochondrial (ND3) gene expression. Our results demonstrated the purity and well distribution of Er 2O3-NPs and revealed that Er2O3-NPs induce strong cytotoxicity on U937 cells, as evidenced by a concentration-dependent reduction in cell viability with an IC50 value of 3.20 µg/ml. Exposure of U937 LCs to the IC50 concentration (3.20 µg/ml) of Er2O3-NPs promoted excessive ROS generation, leading to dramatic damage to genomic DNA and mitochondrial membrane potential, as well as marked dysregulation of apoptotic (p53), anti-apoptotic (Bcl2) and mitochondrial ND3 gene expression. This cascade of events triggered both apoptosis and necrosis in Er2O3-NPs-treated U937 LCs. In conclusion, these fndings highlight the strong in vitro cytotoxic potential of Er2O3-NPs against highly aggressive U937 LCs, mediated by excessive ROS production, which leads to severe genomic DNA and mitochondrial membrane damage, as well as profound alterations in apoptotic, anti-apoptotic and mitochondrial gene expression. Future research is needed to further investigate the potential use of Er2O3-NPs in treating lymphoma and to optimize their therapeutic efcacy
Evaluation of the effect of grape seed extract loaded-chitosan nanoparticles on cryptosporidiosis in dexamethasone immunosuppressed male mice
(Elsevier B.V, 2025-02-25) Karim Tarek; Gehan Safwat; Alyaa Farid
Cryptosporidiosis is a worldwide health problem that results in an economic loss. The disease is caused by the protozoan Cryptosporidium spp. Individuals with suppressed immunity, like those with organ transplantation, cancer and human immunodeficiency virus syndrome, suffer from the infection that may lead to the death. Nitazoxanide (NTZ) is the approved FDA treatment for cryptosporidiosis in immunocompetent individuals. There is an urgent need to find a new natural treatment that can replace NTZ in immunosuppressed hosts. The study aimed to use grape seed extract loaded chitosan nanoparticles (GSEx-CHNPs) in treatment of cryptosporidiosis in immunosuppressed male mice. GSEx was prepared by the alcoholic extraction method followed by the identification of its bioactive components. GSEx-CHNPs were synthesized by ionic gelation method and physically characterized then their activities were examined in vitro. The experimental groups, included immunocompetent and immunosuppressed groups, was treated with NPs for 14 days post infection (PI). The results showed the presence of many phenolic compounds in the GSEx. GSEx-CHNPs significantly improved the loss in animals body weight, cleared the infection and amolerated the serum cytokines levels. GSEx-CHNPs showed anti-cryptosporidial activity especially in immunosuppressed mice model. Where, it amolerated the disturbance in the cytokine profile leading to an anti-inflammatory response.
Calcium titanate nanoparticlesinduced cytotoxicity, genotoxicity and oxidative stress in human nonsmall lung cancer cells
(Nature Publishing Group, 2025-02-21) Hanan R. H. Mohamed; Shahd E. E. Shaheen; Esraa H. Ibrahim; Nesma O. E. Hussein; Gehan Safwat
Calcium titanate nanoparticles (CaTiO3NPs) have garnered significant attention due to their unique properties and excellent biocompatibility, which have led to their increased use in various fields and consumer products. This rise in application necessitates a better understanding of their biological and toxicological effects. However, there is limited data on the cytotoxicity and genotoxicity of CaTiO3NPs in human normal skin fibroblasts (HSF) and non-small lung cancer (A-549) cells. Consequently, this study aimed to explore the effect of 48-hour exposure to CaTiO3NPs on cell viability, genomic DNA integrity, and oxidative stress induction in human cancer A-549 cells, compared to normal HSF cells. The cytotoxicity and genotoxicity of CaTiO3NPs were assessed using the Sulforhodamine B (SRB) cytotoxicity and Alkaline Comet assays, respectively. To estimate possible oxidative stress induction and variation in apoptotic gene expression, reactive oxygen species (ROS) analysis and quantitative real-time polymerase chain reaction (qRT-PCR) were also performed. Our findings demonstrated that exposure to CaTiO3NPs for 48 h resulted in low toxicity toward both normal HSF and cancer A-549 cells, with cell death observed only at high concentrations (100 and 1000 µg/ml). The IC50 value of CaTiO3NPs in both HSF and A-549 cells was greater than 1000 µg/ml; specifically, the IC50 value in A-549 cells at 48 h was 1670.65 µg /ml. However, treatment with CaTiO3NPs for 48 h at the IC50 concentration of 1670.65 µg /ml resulted in significant genomic DNA damage and excessive ROS generation, along with a notable disturbance in the expression level of apoptotic (p53 and Bax) and anti-apoptotic Bcl2 genes in A-549 cells. In contrast, no significant changes were observed in HSF cells treated for 48 h with the same concentration (1670.65 µg /ml) of CaTiO3NPs. Collectively, these findings indicated that despite short-term exposure to CaTiO3NPs causing low cytotoxicity in both normal HSF and A-549 cells. CaTiO3NPs were selectively genotoxic toward A-549 cells. This genotoxicity was mediated through excessive ROS generation, which disrupted genomic DNA integrity and altered the expression of apoptotic genes, triggering apoptosis in A-549 cells. Further in vitro and in vivo studies are needed to fully understand the toxicological and biological properties of CaTiO3NPs.
Purifcation and Immobilization of Burkholderia gladioli Cholesterol Oxidase on Calcium Alginate, with Robust Catalytic Stability for Cholesterol Oxidation In Vitro
(Springer New York, 2025-02-05) Ashraf S. A. El‑Sayed; Ahmed A. Shindia; Walaa M. El‑Badry; Ahmed A. Mostafa; Abdullah A.Al‑Ghanayem; Amgad M. Rady
Cholesterol oxidase (COX) is a key enzyme in diagnostic kits of cardiovascular diseases via oxidation of cholesterol producing smart enantiomerically compounds; however, the enzyme catalytic stability is the challenge. So, the objective of this study was to purify COX from novel endophytic bacterial isolates of medicinal plants that could have unique catalytic efciency for the desired applications. Among the recovered forty bacterial isolates, Burkholderia gladioli EFBL PQ721377, an endophyte of Eruca sativa, had the highest COX productivity (14.7 μmol/mg/min). The COX productivity of B. gladioli has been maximized by with the response surface methodology, giving the highest productivity 30.9 μmol/mg/min, by ~2.0-fold increment compared to control. The enzyme was purifed to its molecular homogeneity with subunit structure 40 kDa. The enzyme was entrapped in Ca-alginate with immobilization yield 87.5%, and the efciency and homogeneity in Ca-alginate beads were assessed by FTIR and SEM–EDX analyses. The free and Ca-alginate-COX conjugates have the same maximum reaction temperature at 37–40 °C, reaction pH at 7.5 and pH stability at 6.5–8.0. The thermal stability of Ca-alginate-COX was increased by ~ 7.0 folds compared to the free one, ensuring the protective role of alginate beads on enzyme tertiary structure. Ca-alginate-COX had a higher potency of oxidation of human serum cholesterol, than the free one, confrming the feasibility of the product release, and allosteric activation of the enzyme, with a reliable operative stability till the ffth cycle, for production of cholest-4-en-3-one, as the precursor of various drugs.
Deciphering the Interplay of the PD-L1/MALT1/miR-200a Axis During Lung Cancer Development
(Wiley-Blackwell, 2025-01-05) Sherien M. El-Daly; Sahar S. Abdelrahman; Mona A. El-Bana; Yasmin Abdel-Latif; Dalia Medhat; Safaa M. Morsy; Hanaa A. Wafay
Lung cancer remains a leading cause of cancer-related mortality worldwide. Our study investigates the involvement of the PD-L1/MALAT1/miR-200a-3p axis in lung tumor progression using a murine model of lung carcinogenesis. Lung tumors were induced in rats, which were divided into groups and sacrificed at different stages of tumor development. A histopathological examination was performed to assess tumor progression. Immunohistochemistry was applied to evaluate the expression of Ki-67 and programmed death-ligand 1 (PD-L1). The level of carcinoembryonic antigen (CEA) and expression analysis of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), miR-200a-3p, and zinc finger E-box binding homeobox 1 (ZEB1) were evaluated for each stage of induction. Immunohistochemical analysis demonstrated a progressive upregulation of the proliferative marker Ki-67 and the immune checkpoint protein PD-L1 during the induction process, indicative of enhanced tumor proliferation and immune evasion. Additionally, CEA levels revealed a progressive increase across induction stages, with a significant increase in advanced tumor stages, highlighting its clinical relevance as a biomarker for lung cancer progression. Expression analysis revealed dynamic upregulation of MALAT1 and downregulation of miR-200a during lung tumor induction, which correlated with advanced tumor stages and elevated PD-L1 expression, suggesting that the negative correlation between MALAT1 and miR-200a is involved in the development of lung tumors. ZEB1 expression exhibited a notable increase in the advanced stages of induction, consistent with its association with aggressive lung cancer. Our findings underscore the interplay between molecular pathways involved in lung tumor development and the potential diagnostic and therapeutic implications of the PD-L1/MALAT1/miR-200a-3p axis.
Egyptian Novel Goose Parvovirus in Immune Organs of Naturally Infected Ducks: Next-Generation Sequencing, Immunohistochemical Signals, and Comparative Analysis of Pathological Changes Using Multiple Correspondence and Hierarchical Clustering Approach
(Multidisciplinary Digital Publishing Institute (MDPI), 2025-01-13) Mohamed A. Lebdah; Amal A. M. Eid; Reham M. ElBakrey; Abd Elgalil. El-Gohary; Mohamed G. Seadawy; Mohamed R. Mousa; Hagar F. Gouda; Nehal I. A. Goda; Mostafa F. El-Hosseny; Ahmed S. El-tahlawy; Rokayya Sami; Rasha A. Al-Eisa; Sarah S. Helal
The present study aims to better understand the nature of currently circulating GPV strains and their pathological impact on the immune system during natural outbreaks among different duck breeds in Egypt. For this purpose, 99 ducks (25 flocks) of different breeds, aged 14–75 days, were clinically examined, and 75 tissue pools from the thymus, bursa of Fabricius, and spleen were submitted for virus detection and identification. Clinical and postmortem findings were suggestive of GPV infection. Concerning the immune system organs, atrophy in the thymus (60.6%), bursa (45.5%), and spleen (38.3%) was the most common gross lesion. Microscopically, the pathological impact of the virus was exhibited by a necrotic thymic cortex with Hassall’s corpuscle disintegration, the disappearance of normal bursal histological morphology accompanied by atrophied follicles and lymphocytic depletion, and apoptosis of B-lymphocytes in lymphoid follicles of the spleen. Furthermore, immunohistochemical examination revealed positive signals of the parvovirus detected in thymic lymphocytes in the cortex, bursa-dependent lymphoid follicle of the medulla, and diffuse positive expression of viral antigens in the spleen. GPV was detected in ducks using polymerase chain reaction, with the highest percentage of positive detection in the bursa of Fabricius (76%). Next-generation sequencing and phylogenetic analysis revealed that the detected virus was a variant of GPV, globally named novel GPV (NGPV), and closely related to Chinese NGPV isolates. To our knowledge, the current study is pioneering to address the immunopathological impact of NGPV among naturally infected ducks confirmed with full genome sequencing and immunohistochemical identification worldwide.

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