OBJECTIVES:Regulation of immune responses in renal transplant recipients is complex, and the CTLA -4 gene plays a crucial role in maintaining immune tolerance. Methylation of CpG regions in the CTLA -4 promoter may affect its expression, potentially influencing transplant outcomes. This pilot study investigated the relationship between CTLA -4 promoter methylation and gene expression in CD8 + T cells of renal transplant recipients. MATERIALS AND METHODS:We analyzed the CTLA -4 promoter methylation profile in CD8 + T cells from 29 renal transplant recipients using bisulfite sequencing. The methylation status of specific CpG regions was correlated with CTLA -4 expression levels to assess any potential influence of methylation on gene regulation. RESULTS:Two patients with hemi -methylated CpG regions in the CTLA -4 promoter showed an increase in CTLA -4 expression. However, no overall correlation between methylation levels and gene expression was observed across the patient cohort. CONCLUSIONS:The findings suggested that immune responses and treatment outcomes may differ considerably between individuals. In addition, other epigenetic mechanisms may play a role in modulating CTLA -4 expression. These results highlight the complexity of immune regulation in renal transplant recipients and the need for further investigation into the epigenetic factors influencing transplant outcomes.

Cyclin-dependent kinase (CDK) inhibitors are promising therapeutic agents for neuroblastoma, particularly in MYCN-amplified tumors; however, the molecular responses associated with CDK inhibition remain incompletely understood. MYCN-amplified BE(2)-C and MYCN-non-amplified SH-SY5Y neuroblastoma cells were treated with Abemaciclib or Arcyriaflavin A. Drug responses were evaluated using MTT, Annexin V ELISA, wound healing, colony formation, quantitative real-time PCR, and Western blot analyses. Exploratory transcriptomic profiling was performed by RNA sequencing. Because only one biological replicate was available per condition, transcriptomic analyses were limited to descriptive expression trends and pathway-oriented interpretation rather than statistical differential expression analysis. Both inhibitors reduced cell viability in a dose- and time-dependent manner, with BE(2)-C cells showing greater sensitivity than SH-SY5Y cells. Treatment also increased Annexin V levels, reduced wound closure, impaired clonogenic growth, and decreased CDK4/CDK6 expression, as confirmed by qRT-PCR and Western blotting. Exploratory transcriptomic profiling identified concordant expression trends in genes associated with cell-cycle regulation, DNA replication, and mitotic progression, with broader pathway modulation observed in MYCN-amplified cells. Overall, CDK inhibition suppresses proliferation-associated phenotypes in neuroblastoma and is accompanied by exploratory transcriptomic changes that identify candidate pathways for future validation and mechanistic investigation of MYCN-associated therapeutic responses.

Purpose Glioblastoma (GBM) is the most aggressive primary brain tumor in adults, characterized by poor prognosis and resistance to standard therapies. Cyclin-dependent kinase (CDK) inhibitors, particularly abemaciclib, have emerged as promising candidates for targeting GBM through cell cycle modulation. However, their molecular mechanisms and interactions with other chemotherapeutics remain to be fully elucidated. Methods In this study, we evaluated the effects of abemaciclib alone and in combination with doxorubicin, cisplatin, or temozolomide (TMZ) on U87 glioblastoma cells. Cell viability (WST-8 assay), wound healing, colony formation, apoptosis (Annexin V ELISA and flow cytometry), and cell cycle progression (PI staining) were assessed. Gene and miRNA expression analyses were also performed to identify mechanistic changes underlying treatment responses. Results Abemaciclib significantly reduced U87 cell viability and enhanced the cytotoxic effects of cisplatin and TMZ in combination treatments. Apoptosis assays revealed increased Annexin V levels, particularly in the abemaciclib-TMZ group. Flow cytometry confirmed G1 phase arrest in abemaciclib-treated cells, while TMZ induced G2/M arrest. Gene expression analysis showed upregulation of pro-apoptotic and cell cycle arrest genes (Casp3, Casp9, p16), and downregulation of CDK4. miRNA profiling demonstrated consistent upregulation of miR-128-3p and modulation of miRNAs linked to GBM progression, including miR-21, miR-34a, and miR-449a-3p. Conclusion Abemaciclib exerts potent anti-glioblastoma activity by modulating cell cycle, apoptosis, and miRNA expression. When combined with cisplatin or TMZ, it enhances therapeutic efficacy in U87 cells. These findings support further investigation of abemaciclib-based combination therapies as a strategy to overcome GBM resistance.

Natural compounds, as honey-derived flavonoids and phenolic compounds, are increasingly investigated for their potential to mitigate skin aging and prevent oxidative stress-induced cellular damages. In this context, a dynamic cell culture model was employed to assess the protective influence of honey pre-treatment on stem cell-associated genes and the Wingless-related integration site (Wnt) signaling pathway following ultraviolet (UV)-induced aging. Using a bioreactor, skin stem cells (SSCs) derived from healthy skin biopsies and human skin fibroblasts (HFF1) were pre-treated with 1% honey for 48 h and then exposed to UV. Real-time quantitative polymerase chain reaction (RT-qPCR) analyses were performed on Wnt signaling and anti-aging molecular responses. Honey pre-treatment enhanced the expression of pluripotency markers (Octamer-binding transcription factor 4 (Oct4); SRY-box transcription factor 2 (Sox2)) and reduced senescence-related cell cycle regulators (cyclin-dependent kinase inhibitor 2A (p16); cyclin-dependent kinase inhibitor 1A (p21); tumor protein 53 (p53)) in SSCs. In UV-damaged SSCs, honey also significantly increased Wnt3a expression. In fibroblasts, honey pre-treatment upregulated Heat shock protein 70 (Hsp70) and Hyaluronan synthase 2 (HAS2) expression, while downregulating caspase-8 (CASP8), indicating a protective role against UV-mediated cellular stress. We also analyzed nitric oxide release and the total antioxidant capacity of cells after treatment. Collectively, these findings suggest that honey may safeguard skin stem cells from UV-induced aging by modulating pluripotency and senescence-associated genes and regulating differentiation through alterations in Wnt signaling. Furthermore, Hsp70 upregulation in fibroblasts appears to strengthen cellular stress responses and support homeostatic stability.

Chronic myeloid leukemia (CML) is a hematological malignancy driven by the BCR::ABL fusion protein. Although tyrosine kinase inhibitors such as imatinib have markedly improved clinical outcomes, treatment resistance remains a major challenge. Natural product-derived compounds may offer alternative therapeutic opportunities. This study investigated the antiproliferative and pro-apoptotic effects of newly synthesized oleanolic acid (OA) derivatives in K562 cells and evaluated their predicted interactions with the ABL kinase domain. OA derivatives were synthesized and characterized by FT-IR and NMR. Cytotoxicity was assessed using the WST-8 assay. Apoptosis, mitochondrial membrane potential, and cell-cycle distribution were analyzed by flow cytometry. Expression levels of BCL2, BAX, Caspase-3, Caspase-8, and Caspase-9 were determined by qPCR, whereas BCR::ABL protein levels were assessed by Western blotting. Boltz-2 complex prediction, molecular dynamics simulations, and MM/PBSA calculations were used to evaluate predicted binding stability, interaction patterns, and energetics. OA-Acid showed the lowest IC₅₀, whereas OACN showed the highest value . Apoptosis was highest in imatinib- and OACN-treated cells, and mitochondrial membrane potential decreased significantly in both groups. OA-Acid and MCM35 produced the greatest G0/G1 accumulation. Apoptosis-related gene expression was altered in a compound-dependent manner. OA-Acid showed the greatest decrease in BCR::ABL protein level. Computational analyses supported conformationally stable predicted interactions of OA, OA-Acid, and OACN within the ABL ATP-binding region. OA derivatives produced distinct cytotoxic, apoptotic, mitochondrial, cell-cycle, and molecular effects in K562 cells. Computational findings supported predicted interactions within the ABL kinase domain, warranting further chemical and mechanistic evaluation.

OBJECTIVES:Hypoxia is a key regulator of renal epithelial cell function and contributes to the pathogenesis of kidney diseases. Although two-dimensional cell cultures are commonly used to study hypoxic responses, three -dimensional spheroid cultures better represent the in vivo microenvironment, including cell -cell and cell -matrix interactions. In this study, we compared hypoxia -induced gene expression in two - and three -dimensional cultures of HK2 cells. MATERIALS AND METHODS:We induced hypoxia at 12 hours in two -dimensional cultures and at 24 hours in three -dimensional spheroids, with confirmation of hypoxia by hypoxia -inducible factor 1α protein expression and vascular endothelial growth factor mRNA upregulation. With quantitative real -time polymerase chain reaction analysis, we noted that long noncoding RNA H19 was significantly upregulated in both culture models, accompanied by modulation of NLRP3, Wnt1, β -catenin, interleukin 1 β, interleukin 6, tumor necrosis factor α, interleukin 10, microRNA -30a -5p, and microRNA -196a. RESULTS:Tumor necrosis factor α and microRNA -30a-5p expression were decreased, with strong interleukin 10 upregulation, in two -dimensional cultures. In three -dimensional cultures, expression levels of both microRNA -30a -5p and microRNA -196a were further upregulated, with less pronounced reduction of tumor necrosis factor α and limited increase of interleukin 10 expression. CONCLUSIONS:Our findings suggested that microRNA -30a -5p and microRNA -196a may play a role in modulating hypoxia -induced cytokine responses, contributing to a more balanced cellular adaptation in three -dimensional cultures. Three-dimensional HK2 spheroids may provide a physiologically relevant model for studying hypoxic stress and may reveal the potential involvement of long noncoding RNA H19 and specific microRNAs in regulation of inflammatory and signaling pathways. Of note, this study represents one of the first investigations of hypoxia -induced gene expression in three -dimensional HK2 cultures, offering a novel contribution to understanding renal epithelial responses under low oxygen conditions.

Purpose: This study aimed to address the limited understanding of complement-associated molecular mechanisms in antibody-mediated rejection (ABMR) by evaluating ABMR-related mRNA and miRNA expression in an in vitro HK-2 cell model exposed to donor-specific anti-HLA antibodies. Materials and Methods: We used the HK-2 cell line derived from normal kidney epithelial cells. An end-stage renal disease patient with HLA class I donor specific antibody, who was registered on the kidney waiting list for a positive serum [DSA(+)] and a healthy male without blood transfusion [negative control DSA (-)] were included in the study. The model includes both complement-dependent cytotoxicity (CDC) with and without complement in cell culture. Then, immune response-related mRNA and miRNA expression levels were evaluated by qRT-PCR. Results: mRNA and miRNA expression levels were analyzed by qRT-PCR, revealing pronounced dysregulation in the DSA(+) CDC C(+) group. Let-7c-5p and miR-155 were markedly upregulated (-103-104-fold), whereas miR-21, miR-217, miR-145, miR-125a-5p, and miR-885 were downregulated (-10-2-10-4-fold). At the mRNA level, strong upregulation of TGF-beta (-50-fold), TNF-alpha (-10-fold), and caspase-3 (-10-fold) was observed, while C1qB was markedly downregulated (-0.1-fold). Additionally, differences in the expression of TNF alpha, C1qB and severalmiRNAs (miR-142-5p, miR-155, miR-125a-5p, miR-21, miR-29b-3p, miR-145, and miR-217) were detected between CDC complement C(-) and C(+) conditions. Conclusion: This study presents the initial characterization of an in vitro ABMR model using kidney tubule epithelial cells and donor specific anti-HLA-A2 (+) serum. Furthermore, this is the first allorecognition model that assesses both C1q mRNA and certain miRNA expressions linked to antibody mediated rejection simultaneously.

Objective: Researchers have established that the fenugreek plant exhibits anti-proliferative effects against various types of cancer, highlighting its potential as a valuable source for the development of new anticancer drugs. This study attempted to assess the anti-cancer effects of fenugreek on the K562 chronic myeloid leukemia (CML) cells due to the Wnt signaling pathway. Materials and Methods: The cytotoxic effect of Fenugreek was performed by 2,3-bis(2-methoxy-4-nitro-5reaction (RT-PCR) to determine gene expression, and western blot was used to detect beta-catenin, c-Jun Nterminal kinase (JNK), calcium/calmodulin-dependent protein kinase (CamK), and protein kinase C (PKC) proteins. Results: The IC50 value of fenugreek extract was 717 g/mL after 24 h, and an in vitro cytotoxic effect was observed on K562 cell lines. JNK, PKC, CamK, and beta-catenin protein levels were reduced in fenugreek extract-treated cells by 81%, 20%, 7%, and 26%, respectively. Conclusion: According to our findings, fenugreek can affect both canonical and non-canonical pathways in K562 cells, particularly via the JNK protein in the planar cell polarity (PCP) pathway. These data corroborates our hypothesis that fenugreek extract possesses adjuvant compounds beneficial for CML treatment.

Neuroblastoma (NB), a common pediatric cancer, is often associated with poor prognosis due to resistance to conventional therapies. Abemaciclib, a selective inhibitor of CDK4/6, is known for its ability to block cell cycle progression and induce cell death in various cancer types. In this study, we explore its potential therapeutic impact on NB by assessing its effects on cell proliferation, apoptosis, and the regulation of microRNAs (miRNAs) that are related to NB progression. Antiproliferative effect of abemaciclib, doxorubicin, cisplatin, and temozolomide (TMZ) were detected by MTT method. Combinations of abemaciclib-doxorubicin, abemaciclib-cisplatin, and abemaciclib-TMZ were also investigated by applying IC50 doses of the drugs for 24 h. ELISA and flow cytometry were performed for apoptosis detection, and for cell cycle analysis, flow cytometry was used. The expression levels of eight apoptosis, threee tumor suppressors, two oncogenes, and nine cell cycle-related genes were analyzed by quantitative PCR. Moreover, the expression levels of five NB-related miRNAs were determined. IC50 doses of abemaciclib, doxorubicin, cisplatin, and TMZ were found to be 4.757, 1.958, 34.21, and 240.7 uM in the 24 h, respectively. The combination of the drugs increased apoptosis and decreased cell migration and colony formation rates. The highest expression level difference was observed in PUMA when control and dose groups were compared. Increased expression levels of hsa-mir-18a-5p and hsa-miR-124-3p were detected in all drug-treated groups compared to the control group. Our results highlight the potential of abemaciclib as a promising treatment strategy for NB, particularly when used in combination with other therapies to overcome resistance and improve clinical outcomes.

Saponaria species are known to contain saponins which have a wide variety of biological activities. But up to now, the phenolic compounds of Saponaria mesogitana have not been clarified. Therefore, this study aimed to determine the phenolic composition and some biological activities of S. mesogitana for the first time. The antioxidant activities of the methanol and water extracts were assessed using the DPPH, FRAP and beta-carotene/linoleic acid assays, while the total secondary metabolite content, including phenolics, flavonoids and saponins, was also determined for both extracts. Based on the antioxidant activity and total phenolic and flavonoid contents, further HPLC analysis, as well as anticancer and antimicrobial activity experiments, were conducted using the methanol extract. The anticancer potential was assessed using the MTT assay and wound healing migration test, while antibacterial activity was evaluated through disc diffusion and MIC assays. Additionally, the antibiofilm properties of the extract were examined using the crystal violet method. The methanolic extract showed high antioxidant activity, while caffeic acid and epicatechin were characterized as major phenolic compounds by HPLC. S. mesogitana inhibited not only bacterial growth but also the levels of migration of SHSY-5Y cancer cells. These findings indicate that S. mesogitana possesses potent antioxidant, anticancer, antimicrobial and antibiofilm activities associated with its bioactive phenolic constituents.

Fibromyalgia syndrome (FMS), affecting 3–10

Purpose: Honey is increasingly recognized for its medicinal and nutritional benefits, including its antibacterial, anti-inflammatory, antioxidant, anti-ulcer, and anti-tumor properties. This study aimed to evaluate the effects of Anatolian honey (Southeast Anatolia) on the expression of pro-apoptotic genes and chemotactic molecules in breast cancer cells. Materials and Methods: The IC50 concentration of Anatolian honey, determined to be 2.7% (w/v) at 48 hours, was used for treatment. Gene expression analysis was performed using the SYBR Green quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique. Results: The expression levels of the pro-apoptotic genes Bax and Caspase-3 increased by 6.05-fold and 2.01-fold, respectively, while the chemotactic molecule CCL2 exhibited a 6.45-fold upregulation. In contrast, CCL5 and CX3CR1 showed minimal changes in expression, with fold changes of 1.04 and 1.11, respectively. Anatolian honey promoted apoptosis by increasing Bax and Cas3 expression. Additionally, the elevated CCL2 expression suggests a potential role in recruiting monocytes and facilitating their differentiation into macrophages within the tumor microenvironment. Conclusion: Anatolian honey exhibits pro-apoptotic and immunomodulatory effects in breast cancer cells by upregulating Bax, Cas3, and CCL2 expression. These findings highlight its potential as a complementary therapeutic agent.

Objective: Glioblastoma multiforme is one of the most aggressive brain tumors, with a low survival rate and limited treatment options. Moreover, resistance to chemotherapy complicates the treatment. Plant extracts are commonly used to overcome drug resistance and support currently used chemotherapies. This study aimed to determine the anticancer effect of fenugreek and cinnamon extracts in combination with doxorubicin and cisplatin on U87 glioblastoma cells. Methods: Fenugreek and cinnamon extracts were donated by Indus Biotech (India). The 50% viability of fenugreek, cinnamon, doxorubicin, and cisplatin was determined by MTT assay. The combination of plant extracts and drugs was also applied to U87 cells. Apoptosis and cell cycle analysis were performed by flow cytometry. Results: IC50 concentrations for 24h were found to be 224.8 µg/mL for fenugreek, 3269 µg/mL for cinnamon, 3.2 µM for doxorubicin, and 770 µM for cisplatin, respectively. Fenugreek, doxorubicin, and fenugreek-cisplatin groups were found to be more cytotoxic than cinnamon-doxorubicin and cinnamon-cisplatin groups. The treatment of plant extracts and drugs seems to shift the cell cycle distribution, particularly increasing the G0/G1 phase. Conclusion: These results are thought to be meaningful in understanding the potential effectiveness of the combination of fenugreek and cinnamon with traditional chemotherapeutic agents in cancer treatment.

Objective: After kidney transplantation, CD8+ T cells can infiltrate the kidney and cause necrosis, tubulitis, and even transplant rejection. For this reason, control of the T cell response is very important, and T cell immunoglobulin and mucin domain 3 (TIM-3) and programmed death 1 (PD-1) molecules play a role in regulating the T cell response. It is thought that the levels of TIM-3 and PD-1 expressions may be guiding in determining the clinical course after transplantation. This study aimed to determine the relationship between the mRNA levels of PD-1 and TIM-3 genes in peripheral blood samples taken from kidney transplant patients and the clinical conditions of the patients. Material and Method: 60 peripheral blood samples were collected from 30 kidney transplant patients, both pre-transplantation (pre-tx) and post-transplantation (post-tx). CD8+ T cells were separated from other lymphocytes by magnetic cell separation system (MACS) and their purity was determined by flow cytometry. Then, RNA was isolated and after cDNA conversion, the expressions of PD-1 and TIM-3 genes were determined by real-time polymerase chain reaction. Results: While it was determined that the TIM-3 gene expression level increased in patients with acute tubular necrosis, antibody-mediated rejection and cell-mediated rejection findings (p

Wnt-signaling pathway plays a crucial role in the pathogenesis and progression of Chronic Myeloid Leukemia (CML). sFRP1 is involved in the suppression of the Wnt-signaling pathway and has been shown to be epigenetically silenced by promoter hypermethylation during CML progression. DNMT3A plays a crucial role in promoter hypermethylation and is responsible for establishing methylation patterns. We aimed to analyze the relationship between sFRP1 expression and DNMT3A, TET1, TET2 and TET3 proteins that are responsible for maintaining cellular methylation patterns; along with miRNAs miR144-3p and miR-767-5p that are known to be associated with these proteins. CML cell lines K562 and K562S which stably expresses sFRP1, were used to compare the changes in miR144-3p and miR-767-5p expression. DNMT3A, TET1, TET2 and TET3 protein levels were analyzed by Western blot. In K562S cells the expression of miR-144-3p and miR-767-5p were decreased along with DNMT3A and TET1 protein levels. On the contrary, TET2 protein was increased. Our results support other reports involving sFRP1 and methylation dynamics; as well as opening new avenues of exploration. Our data supports the conclusion that re-expression of sFRP1 protein alters the expression of factors that play important roles in the overall methylation patterns in the leukemic cell line K562.

Nanostructured materials used have unique properties and many uses in nanotechnology. The most striking of these is using herbal compounds for the green synthesis of nanoparticles. Among the nanoparticle types used for green synthesis, gold nanoparticles (AuNPs) are used for cancer therapy due to their stable structure and non-cytotoxic. Lung cancer is the most common and most dangerous cancer worldwide in terms of survival and prognosis. In this study, Nasturtium officinale (L.) extract (NO), which contains biomolecules with antioxidant and anticancer effects, was used to biosynthesize AuNPs, and after their characterization, the effect of the green-synthesized AuNPs against lung cancer was evaluated in vitro. Ultraviolet‒visible (UV‒Vis) spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), multiple analysis platform (MAP), and Fourier transform infrared (FT-IR) spectroscopy analyses were performed to characterize the AuNPs prepared from the N. officinale plant extract. Moreover, the antioxidant activity, total phenolic and flavonoid contents and DNA interactions were examined. Additionally, A549 lung cancer cells were treated with 2–48 µg/mL Nasturtium officinale gold nanoparticles (NOAuNPs) for 24 and 48 h to determine the effects on cell viability. The toxicity of the synthesized NOAuNPs to lung cancer cells was determined by the 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide (MTT) assay, and the anticancer effect of the NOAuNPs was evaluated by apoptosis and cell cycle analyses using flow cytometry. The average size of the NPs was 56.4 nm. The intensities of the Au peaks from EDS analysis indicated that the AuNPs were synthesized successfully. Moreover, the in vitro antioxidant activities of the NO and NOAuNPs were evaluated; these materials gave values of 31.78 ± 1.71

Aim: The goal of this study was to investigate the expression levels of microRNAs (miRNAs) (miR-196b, miR-10a, miR-31-5p, and miR-338-3p) which regulate the genes involved in the proliferation and function of cells functioning in the inflammatory processes in Celiac patients' blood and tissue samples. Celiac disease (CD) is an inflammatory disease which affects people who are genetically predisposed to gluten consumption. The only treatment for this disease is a gluten-free diet. Materials and Methods: The miRNA expressions were determined in blood and tissue samples from 12 pediatric patients with CD and from 8 healthy children using quantitative real-time PCR (qRT-PCR) and SybrGreen dye. The gene expression levels of miRNAs such as miR-196b, miR-10a, miR-338-3p, and miR-31-5p were compared between the two groups. Results: There was a significant difference only in miR-10a gene expression levels between the control and patient blood samples. The greatest difference between the tissue and blood samples within the CD group were found in the expressions of miR-31-5p and miR-338-3p. It was seen that the patients’ human leukocyte antigen tissue type was not associated with their miRNA expression profiles. In addition, there was no significant correlation between their Marsh classification and gene expression levels. Conclusion: The significantly low level of miR-10a may be related to CD due to its effect on the immune response. Additionally, miR-10a may have potential as a non-invasive biomarker in the diagnosis of CD.

recognized since the 1970s that macrophages play a role in graft rejection (3) .Macrophages play an essential role in host defense, inflammatory processes, ischemia-reperfusion injury, and tissue homeostasis (4) .In addition, they are involved in ÖzOrgan nakli, son dönem organ yetmezliği olan hastalar için hayat kurtaran bir tedavi seçeneğidir.Greft reddi, organ naklinden sonra gelişebilecek ciddi bir komplikasyondur ve patofizyolojisi birçok değişkene bağlıdır.Makrofajlar, doğuştan gelen bağışıklık sisteminin temel hücre gruplarındandır.Klinik çalışmalar, makrofajların antijenleri tanıdığını ve greft reddinde önemli rol oynadığını göstermiştir.Makrofaj infiltrasyonu, artan greft reddi insidansı ile ilişkilidir.Uzun ve kısa süreli greft reddini engellemek ve greft sağkalımını artırmak için makrofaj hedefli terapötik çalışmalara ihtiyaç vardır

Objective: Celiac disease (CD) is one of the most common autoimmune disorders in which gluten damages the small intestine. The CTLA-4 and FOXP3 genes play an important role in immune tolerance, so it is hypothesized that polymorphisms of these genes may be related to celiac disease. Our study aimed to investigate the associated with celiac disease and the CTLA-4 +49 A/G (rs231775) and FOXP3 -3279 C/A (rs3761548) polymorphisms by comparing celiac disease patients with a healthy control group. Material and Methods: The single nucleotide polymorphisms (SNP) of +49 A/G in CTLA-4 (rs231775) gene and -3279 C/A in FOXP3 (rs3761548) gene were studied by Polymerase Chain Reaction- Restriction Fragment Length Polymorphism (PCR-RFLP) method in 125 pediatric celiac patients and 100 healthy controls. Results: The A and G alleles of the CTLA-4 gene were found more frequently in the celiac patient group than in the control group. In addition, the A and C alleles of the FOXP3 gene were found more frequently in celiac disease patients than in healthy controls. There were no statistically significant results for the two polymorphisms CTLA-4 +49 A/G and FOXP3 -3279 C/A based on genotype or allele frequency (p > 0.05). When analyzing the risk allele, the FOXP3 gene polymorphism -3279 C/A proved to be significant in CD patients (p

Background: Multiple drug-delivery systems obtained by loading nanoparticles (NPs) with different drugs that have different physicochemical properties present a promising strategy to achieve synergistic effects between drugs or overcome undesired effects. This study aims to develop a new NP by loading quercetin (Que) and valproic acid (VPA) into chitosan. In this context, our study investigated the antioxidant activities of chitosan NPs loaded with single and dual drugs containing Que against oxidative stress. Method: The synthesis of chitosan NPs loaded with a single (Que or VPA) and dual drug (Que and VPA), the characterization of the NPs, the conducting of in vitro antioxidant activity studies, and the analysis of the cytotoxicity and antioxidant activity of the NPs in human neuroblastoma SH-SY5Y cell lines were performed. Result: The NP applications that protected cell viability to the greatest extent against H2O2-induced cell damage were, in order, 96 µg/mL of Que-loaded chitosan NP (77.30%, 48 h), 2 µg/mL of VPA-loaded chitosan NP (70.06%, 24 h), 96 µg/mL of blank chitosan NP (68.31%, 48 h), and 2 µg/mL of Que- and VPA-loaded chitosan NP (66.03%, 24 h). Conclusion: Our study establishes a successful paradigm for developing drug-loaded NPs with a uniform and homogeneous distribution of drugs into NPs. Chitosan NPs loaded with both single and dual drugs possessing antioxidant activity were successfully developed. The capability of chitosan NPs developed at the nanometer scale to sustain cell viability in SH-SY5Y cell lines implies the potential of intranasal administration of chitosan NPs for future studies, offering protective effects in central nervous system diseases.