In aqueous media, enzyme–ligand recognition is shaped not only by direct contacts but also by solvent-mediated interaction networks that persist across time scales. Here, cholinesterases (AChE and BChE) and cytosolic human carbonic anhydrases (hCA I and hCA II) were investigated as two disease-relevant enzyme systems in solution, and twelve thiourea–pyrimidine benzenesulfonamide derivatives were synthesized and structurally characterized. Enzyme inhibition was quantified by steady-state kinetics; inhibition constants (KI) were obtained by nonlinear global fitting of untransformed rate data, with Lineweaver–Burk plots used only for visualization. To connect solution-phase inhibition with molecular recognition, docking was validated by redocking (heavy-atom RMSD <2.0 Å across targets), and representative complexes were examined by 500 ns explicit-solvent MD simulations (TIP3P, 0.15 M NaCl, NPT), revealing stable binding modes and persistent, frequently water-mediated interaction patterns consistent with experimental trends. Cellular relevance was assessed in MKN-28 gastric cancer cells with L929 fibroblasts as a non-malignant control. Compounds with improved cytotoxic selectivity toward MKN-28 were identified, and the lead compound (11) was further evaluated by Annexin V/PI flow cytometry, showing a shift from viable cells toward apoptotic populations under the tested conditions. In silico ADMET profiling suggested moderate lipophilicity across the series, while highlighting elevated polar surface area and compound-dependent toxicity flags as parameters to monitor during optimization. Overall, thiourea–pyrimidine benzenesulfonamides emerge as multifunctional modulators of ChEs and cytosolic hCAs within the applied biochemical and cellular assays.

Glioblastoma (GBM) is one of the most aggressive and lethal primary brain tumors in adults, with limited therapeutic options, highlighting the need for new molecular targets and therapeutic agents. This study describes the synthesis and characterization of two novel series of thiazole-linked hydrazone derivatives (3a-g and 4a-g) and their evaluation as potential human NAD(P)H:quinone oxidoreductase 1 (NQO1) inhibitors for GBM treatment. Cytotoxicity assays against U87 glioblastoma and L929 normal mouse fibroblast cells identified compounds 3e (IC50 = 9.95 +/- 1.08 & micro;M) and 4d (IC50 = 3.27 +/- 0.62 & micro;M) as lead molecules with high selectivity indices. Both compounds effectively inhibited NQO1 activity and increased total reactive oxygen species (ROS) levels. Treatment with 3e and 4d induced mitochondrial-mediated apoptosis by elevating Bax expression and reducing Bcl-2 expression, resulting in a higher Bax/Bcl-2 ratio. Molecular docking studies showed that 3e (-9.0 kcal/mol) and 4d (-7.3 kcal/mol) exhibited strong binding affinities for the NQO1 active site, similar to dicoumarol. These findings suggest that 2-thiazolyl hydrazone analogues represent promising leads for further optimization as novel NQO1-targeting agents for GBM.

Bisphenol A (BPA) is an endocrine-disrupting chemical known to cause testicular toxicity through oxidative stress and apoptosis. Panax ginseng (PG) is a natural product with anti-inflammatory effects. This study aimed to evaluate the protective effect of PG against BPA-induced testicular damage in rats. Thirty-two Wistar Albino rats aged 10-12 weeks were divided into four groups: Control, PG, BPA, and BPA + PG. BPA (50 mg/kg/day) and PG (100 mg/kg/day) were orally administered for 6 weeks. BPA significantly increased serum total oxidant status and decreased antioxidant status (p < 0.001, p < 0.001). Also, the levels of superoxide dismutase (an antioxidant enzyme) (p = 0.0005) and androgen receptor-mRNA (an androgen signaling marker) decreased (p = 0.014). However, caspase 3 (an apoptosis marker) (p = 0.0067), 8-hydroxy-2'-deoxyguanosine (a marker of oxidative DNA damage) (p < 0.001), and tumor necrosis factor-α (a proinflammatory cytokine) (p < 0.001) levels increased in testicular tissues. Rats treated with PG showed improvements in all oxidative and inflammatory markers and significantly restored androgen receptor expression. Histopathological examination revealed degeneration in seminiferous tubules and reduced spermatozoa in the BPA group, while the BPA + PG group showed marked improvement. These findings suggest that PG may alleviate oxidative stress-related testicular damage at both molecular and histological levels and may offer insights for future clinical studies.

Hepatic encephalopathy (HE), complication of liver dysfunction, leads to neurocognitive impairments. Artichoke (Cynara scolymus L.) has been traditionally used for its antioxidant, anti-inflammatory and hepatoprotective properties. This study evaluates artichoke leaf and receptaculum extracts in cholestasis and HE in a rat model. Wistar rats were divided into 6 groups: sham-control, bile duct ligation (BDL), and BDL with low/high-dose leaf or receptaculum extracts. After BDL, physiological saline and extracts (250/500 mg/kg) were administered orally for 28 days. Cognitive activity was evaluated using Morris water maze and novel object recognition tests on day 28. Artichoke extract regulated liver enzymes and bilirubin at high-doses and significantly increased antioxidant enzyme activities reduced by BDL. Elevated 8-Hydroxyguanosine (8-OHdG) levels decreased in liver and brain tissues. Similarly, artichoke extracts reduced cytokine and hydroxyproline (HP) levels elevated by cholestasis. Following BDL, Na⁺/K⁺-ATPase levels in brain and liver tissues decreased, while artichoke extract reversed this. Artichoke, particularly high-dose receptaculum, improved impaired performance and increased time in the target quadrant after BDL. Both artichoke leaf and receptaculum extracts improved recognition. Artichoke treatments, especially high-dose receptaculum, reduced hepatic and neuronal damage and improved histological appearance. These findings highlight the therapeutic potential of artichoke extracts for liver fibrosis and related neurocognitive disorders.

Background/Objectives: Burns are a prevalent health concern that manifest on the skin’s surface or within organs due to various traumas and necessitate prompt intervention. The healing process of the skin involves a sequence of time-dependent events, commencing with the activation of growth factors and culminating in the expression of various genes. To expedite the healing process of burn wounds, there is a need to develop biodegradable materials and new technologies that are compatible with the skin. Methods: In this study, the roles of tilapia (TL, Oreochromis niloticus) fish skin in burn wound treatment processes were investigated. TL or TL-alginate hydrogels (AGTL) were applied to a burn wound created in Sprague Dawley rats for 7 and 14 days. Following the administration of treatment, the levels of hydroxyproline, a critical element in tissue reorganization, along with the gene expression levels of COL1A1, COL3A1, MMP-2, and MMP-9, and the protein expression levels of MMP-2 and MMP-9 were evaluated. Results: Wound closure processes were faster in AGTL-groups compared to TL-groups, and hydroxyproline levels were found to be higher. While the increase in MMP-2 levels was less, the increase in MMP-9 gene and protein levels was greater in the AGTL-group. Concurrently, COL1A1 levels decreased over 14 days, while COL3A1 levels increased in the AGTL-group. Conclusions: Consequently, it was determined that the biological substances in the TL structure, in conjunction with alginate, were effective in the healing and reorganization of the wound tissue. This finding suggests that tilapia may provide a valuable source of insights for future studies aimed at developing effective wound dressings for wound tissues.

Cisplatin (Cis) is the first-line chemotherapy for treating the non-small-cell lung cancer. However, its low solubility, low bioavailability, and potential side effects limit its use. To overcome these drawbacks, novel pH-sensitive liposomal Cis formulations have been developed using a rapid and practical ethanol injection technique. In this study, two different liposome types were prepared, one based on phosphatidylcholine and cholesteryl hemisuccinate (CHEMS), the other containing 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) in combination with CHEMS. Their homogeneity, smaller particle sizes (< 200 nm) and drug integration were confirmed using the DLS method and FT-IR and SEM-EDS, respectively. The loading efficiency was changed from 35 to 50% depending on the composition. In vitro drug release studies showed a minimum release at physiological pH (7.4) and a significantly increased release at acidic pH (5.5), in particularly for DOPE liposomes, indicating the higher pH-sensitivity. Cytotoxicity analyses performed in A549 lung cancer cell line showed that both liposomal formulations exhibited stronger antitumour effects compared to free Cis. This effect was supported by increased apoptotic activity confirmed by Annexin-V/PI staining method. These findings suggest that the pH-sensitive liposomes developed in this study offer a promising and scalable approach to enhance the selective delivery and therapeutic efficacy of Cis.

New, selective and effective anticancer agents are urgently needed in drug research and development studies. For this purpose, some new 2,4-dihydro-3H-1,2,4-triazole-3-thione derivatives based on clopidogrel were synthesized and characterized using spectral techniques, including IR, 1H NMR, 13C NMR and MS. The anticancer activities of the synthesized compounds were evaluated in vitro against human prostate cancer cells (PC3) and human umbilical vein endothelial cells (HUVEC). Among these compounds, the ethyl chain-bearing compound 3 and the 4-methoxyphenyl-bearing compound 8 exhibited the most selective anticancer activities against PC3 with IC50 values of 6.81+2.14 mu M and 4.68+1.76 mu M, respectively. Apoptotic cells were investigated with AO/EB staining. Total DNMT enzyme activity and protein expression levels (DNMT1, DNMT3a, and DNMT3b) were also analyzed in compound 3 and compound 8 treated cells. Compounds 3 and 8 showed DNA methyltransferase inhibitory activity of 63.02+1.40% and 54.08+6.35%, respectively. Molecular docking studies were performed to elucidate the interaction of compounds 3 and 8 with the DNMT1 enzyme. As a result, 3 and 8 were reported to be candidates compounds for the development of DNA methyltransferase inhibitors with potent anticancer activity.

Building upon previous research on hydrazone-bearing aryloxyacetic acid derivatives, a novel series of 5-nitroguaiacol-based hydrazones (3a-l) was synthesized and characterized spectroscopically. Their in vitro cytotoxic effects were evaluated against human non-small cell lung cancer (A549) and normal bronchial epithelial cell lines (BEAS-2B) using the MTT assay. Compounds 3d and 3k exhibited selective and potent cytotoxicity (IC50 = 10.24 mu M and 4.99 mu M, respectively) and strong aldolase A (ALDOA) inhibition at 10 mu M (89.89% and 75.19%). Further mechanistic studies revealed that both compounds decreased HIF-1 alpha expression and modulated apoptotic pathways by upregulating Bax and downregulating Bcl-2 protein levels. Molecular docking studies supported the experimental findings, demonstrating significant interactions of 3d and 3k with key residues in ALDOA, Bax, and Bcl-2, suggesting a dual mechanism involving glycolysis inhibition and apoptosis induction. These results indicate that 3d and 3k are promising lead compounds for targeted lung cancer therapy.

Although the incidence of Alzheimer's disease increases with age, the number of effective drugs in the fight against this disease remains insufficient. In this regard, a new series of hydrazide-hydrazone derivative compounds ( 3a - 3n ) was synthesized and their structures were elucidated using spectral techniques. Then, all compounds were tested for their in vitro antioxidant and anticholinesterase activities. Compound 3i was found to have the highest antioxidant activity in the series with 63.750 ± 0.033 µM and 44.210 ± 0.058 µM SC 50 values in the DPPH and ABTS methods, respectively. Compound 3i exhibited significantly higher inhibitory properties than the reference standard donepezil with IC 50 values of 1.850 ± 0.013 µM and 3.680 ± 0.034 µM against AChE and BChE enzymes, respectively. The cytotoxicity and AChE inhibition potential of the compounds on the SH-SY5Y cell line were also evaluated. Compounds 3i and 3l were found to have the highest AChE inhibition (81.03 ± 2.05% and 83.84 ± 2.46%) in SH-SY5Y cells, respectively. Compound 3l also maintained cell viability at 100 µM concentration. The most active compounds in the series were investigated as competitive or noncompetitive inhibitors against AChE and BChE by enzyme kinetic studies. Moreover, molecular docking and MD simulation studies were used to describe the enzyme-ligand interactions and their stability.

Objective: To evaluate the thiol/disulphide homeostasis and level of vascular endothelial growth factor (VEGF) in saliva of children with type 1 diabetes mellitus and gingivitis. Methodology: Forty children with type 1 diabetes mellitus [DM] and 40 systemically healthy [H]) children were recruited for the study. Based on children’ periodontal and systemic health status, they were divided into four subgroups: 1) systemically and periodontally healthy children (Hh), 2) systemically healthy children with gingivitis (Hg), 3) diabetic children with periodontal health (DM+h), and 4) diabetic children with gingivitis (DM+g). Periodontal clinical parameters were recorded. A new automated technique was used to measure the thiol/disulphide homeostasis parameters, and ELISA was used to measure the amounts of VEGF in saliva. Results: Children with or without diabetes had comparable clinical periodontal parameters and salivary VEGF levels (p>0.05). GI, PI, PD, and disulphide levels were significantly higher in the gingivitis subgroups than in the periodontally healthy subgroups (p<0.001). The gingivitis subgroups had significantly higher amounts of VEGF compared to the periodontally healthy subgroups (p<0.001). Conclusions: Thiol/disulphide homeostasis shifts towards disulphide direction in diabetic children with gingivitis. Thiol/disulphide homeostasis and salivary VEGF levels may be diagnostic markers of gingival inflammation.

Objective: Ovarian cancer has the highest mortality rate in women and it has a poor response rate to treatment due to its late diagnosis and is frequently resistant to currently used cisplatin-based treatment methods. Astragaloside IV (As-IV), a bioactive compound and natural tripeptide glycoside known as an antioxidant, has drawn attention in Chinese medicine for its healing properties. Many studies have shown that it has anti-inflammatory, antidiabetic, antitumoral, and anti-angiogenic properties.Materials and Methods: In our study, we first rendered ovarian cancer cells (OVCAR-3) resistant to cisplatin and then applied determined doses of As-IV (40 μg/mL) and (70 μg/mL) to OVCAR-3 cells and cisplatin-resistant ovarian cancer cells (OVCAR-3- CisR). The cell viability capacity, variation of BAX/BCL-2 gene expression, and regulation of the SDF-1/CXCR4 chemokine axis protein and their gene expressions were investigated.Results: According to the findings, As-IV administration suppressed metastasis by lowering the colony formation potential of cisplatin-resistant ovarian cancer and down-regulating the SDF-1/CXCR4 axis, and increasing the ratio of BAX/BCL-2 mRNA and protein levels due to BAX up-regulation and BCL-2 down-regulation.Conclusion: As a result, we showed that As-IV, used as an antioxidant, can be used as an effective anticancer agent to improve response to the currently used cisplatin-based treatment in cases of drug resistance in ovarian cancer.

Designing new compounds from existing chemotherapeutic drugs to enhance inhibitory effects on tumor cells while overcoming multidrug resistance is one of the important strategies for new drug discovery in medicinal chemistry. A new series of urea and thiourea derivatives based on Lenalidomide as potential anticancer agents have been designed and synthesized. In vitro anticancer activity assay against Caki cancer cells and HUVEC endothelial cells revealed that 1-(4-methylphenyl)-3-[2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl]urea (11) exhibited the highest anticancer activity and selectivity in the series with IC50 values of 9.88 and 179.03 mu M, respectively. Among the compounds, 11 showed significant HDAC1 inhibiton of 68.02 +/- 2.44% at 10 mu M concentration. TGF-beta, Bax, Bcl-2 protein levels and scratch assay were analyzed in Caki cells. As a result, compound 11 induced apoptosis in Caki cells. In this study, it has been demonstrated that compound 11 can be a lead compound for further detailed investigation in renal cancer treatment. Through molecular docking studies, it was determined that the most active compound, 11, forms stable interactions with key residues in the enzyme's active site, particularly engaging in hydrogen bonds with GLY149 and coordinating with the zinc ion in the HDAC1 active site. These interactions are crucial for the observed inhibitory activity. Molecular dynamics simulation revealed the binding event of the most active compound with class I histone deacetylase and the stability of the complex in a biological environment.

In an effort to develop new and effective therapeutic agents for Alzheimer's disease, a series of hydrazone derivatives bearing piperidine rings have been designed and synthesized. The chemical structures of the compounds were characterized by various spectroscopic techniques. In vitro antioxidant and cholinesterase activities of the compounds were evaluated. Among the compounds, N12 exhibited the most antioxidant activity in all methods (CUPRAC, FRAP, DPPH, ABTS). In vitro acetylcholinesterase (AChE) activity results of the compounds showed good IC50 values between 14.124 ± 0.084 and 49.680 ± 0.110 µM were obtained (IC50 = 38.842 ± 0.053 µM for Donepezil). Among the compounds, N7 and N6 are much more effective derivatives than the standard compound donepezil with IC50 values of 14.124 ± 0.084 and 17.968 ± 0.072 µM, respectively. In vitro, butyrylcholinesterase (BChE) inhibition values of the compounds were between 13.505 ± 0.025 and 52.230 ± 0.027 μm. Among the compounds, N6 has the highest BChE inhibition with an IC50 value of 13.505 μm in the series. The cytotoxicity and AChE inhibitory activity of the compounds on SH-SY5Y cell lines were also evaluated. Kinetic studies were also performed to determine the behavior of the compounds as competitive or noncompetitive inhibitors. The binding modes of N6, which was determined to be highly effective according to in vitro analyses, with AChE and BChE were investigated using molecular docking studies, and the stability of the complexes was determined by molecular dynamics simulations. These findings indicated that AChE and BChE enzymes maintained their overall structural stability and compactness during interactions with compound N6.

In this study, synthesis of some new hydrazone derivatives based on articaine was carried out. MDA-MB231 (triple negative human breast cancer cells) and HUVEC (human umbilical vein endothelial cells) cells were used to investigate the cytotoxic activity of hydrazone compounds. Induction of apoptosis and cell viability were assessed by AnnexinV-PI binding levels and Bax-Bcl2 gene expression levels. Inhibitory activities of the compounds against carbonic anhydrase enzyme were also evaluated. Compounds 2b and 2m exhibited the highest cytotoxic activity against MDAMB-231 cells with IC50 values of 16.62 +/- 1.18 mu M and 18.56 +/- 2.36 mu M, respectively. Similarly, the CA inhibition of 2b and 2m was also determined to be the highest in the series.

Objective: Nuclear factor kappa B (NF-κB) is one pathway that controls the expression of genes involved in many cancer events such as proliferation, apoptosis, metastasis, and invasion. Usnic acid is a molecule with many biological effects such as being anticholinergic, gastroprotective, anti-inflammatory, anti-cancerous, and especially antioxidant. This study aims to mechanistically examine the apoptotic behaviors of usnic acid in DU145 prostate cancer cells and the molecules it acts on in the NF-κB pathway. Materials and Methods: This study investigates the apoptotic changes in DU145 cells after usnic acid administration through JC-1 staining and caspase-3 activity measurements. In addition, it tests the effects of usnic acid on subunit p50 and p65 protein and gene expressions in the NF-κB pathway through the respective Western blot and qPCR measurements. Results: The IC50 values of usnic acid at 24 and 48 h in DU145 cells were calculated as 167.06±12.35 μM and 42.15±3.76 μM, respectively. In addition, JC-1 staining showed usnic acid-treated DU145 cells to trigger apoptosis by increasing the membrane permeability of their mitochondria. NF-κB p50 protein expression was also found to be suppressed after usnic acid administration. Conclusion: The results of this study show usnic acid administration to suppress proliferation and to induce mitochondrial apoptosis by suppressing the NF-κB pathway in DU145 cells. This effect of usnic acid indicates it to be combinable with chemotherapeutic agents and evaluable as an alternative in cancer treatment.

In this study, the effect of applying wharton jelly mesenchymal stromal cells (WJ-MSC) isolated from the human umbilical cord tissue on the neonatal mouse model caused experimental asphyxia in mice was investigated. WJ-MSC surface markers (CD44, CD90, CD105) were characterised by immunofluorescence staining, and pluripotency genes (Nanog, Oct-4, Sox-2) were characterised by qPCR. Blood, prefrontal cortex, cerebellum, hippocampus, lung, heart, kidney, and liver tissues were analysed twenty days after subcutaneously administered WJ-MSC. WJ-MSC administration significantly decreased serum TNF-α, NSE, GFAP, and IL-6 levels in the asphyxia mice. It was determined that WJ-MSC application in tissues accelerated cell regeneration and decreased oxidative stress. In conclusion, this study showed that multiorgan damage in asphyxia could be prevented by applying WJ-MSC at an early stage. Therefore, WJ-MSC application in infants with neonatal asphyxia in the clinic may be an innovative method in the future.

Organoid models have gained importance in recent years in determining the toxic effects of drugs in cancer studies. Organoid designs with the same standardized size and cellular structures are desired for drug tests. The field of microfluidics offers numerous advantages to enable well-controlled and contamination-free biomedical research. In this study, simple and low-cost microfluidic devices were designed and fabricated to develop an organoid model for drug testing for renal cancers. Caki human renal cancer cells and mesenchymal stem cells isolated from human umbilical cord were placed into alginate hydrogels. The microfluidic system was implemented to form size-controllable organoids within alginate hydrogels. Alginate capsules of uniform sizes formed in the microfluidic system were kept in cell culture for 21 days, and their organoid development was studied with calcein staining. Cisplatin was used as a standard chemotherapeutic, and organoid sphere structures were examined as a function of time with an MTT assay. HIF-1α, CXCR4 and CXCL-12 chemokine protein, and CXCR4 and CXCL-12 gene levels were tested in organoids and cisplatin responses. In conclusion, it was found that the standard renal cancer organoids made on a lab-on-a-chip system can be used to measure drug effects and tumor microenvironment responses.

Mesenchymal stem cells can be obtained and multiplied from various sources and have a very high capacity to release exosomes. Exosomes are nano-sized extracellular vesicles containing biological signaling molecules. This study aimed to determine the effect of MSC-derived exosomes as a drug delivery system for paclitaxel in cervical cancer cells. In this study, human MSC were isolated from wharton jelly of umbilical cord tissue (WJ-MSC), and cells were characterized by CD44, CD90, CD105, and CD34 staining. Exosomes were released in WJ-MSC cells with serum-starved conditions for 48 hours, and particle sizes and structures were examined with zeta-sizer and TEM. In addition, exosomes CD9, CD63, and CD81 markers were checked by western blot. Paclitaxel was loaded into exosomes (Exo-PAC) by electroporation and then incubated with Hela cervical cancer cells for 24 hours. TGF-β, SMAD, Snail, Slug, β-catenin, Notch, Caspase-3, Caspase-9, Bax, Bcl-2 protein and gene expression levels were analyzed in Hela cells. As a result, low concentration Exo-PAC induced apoptosis, and suppressed epithelial-mesenchymal transition proteins in Hela cells. In this study, it has been demonstrated that WJ-MSCs can be used as drug delivery systems for cervical cancer if exosomes are produced scalably in the future.

Exosomes have come to the fore in drug delivery systems due to their biological-based and immune-suppressing properties. In this study, we investigated the effect of doxorubicin loading of exosomes isolated from human platelets on breast cancer. Exosomes released from ADP (1 µM)-activated platelets were isolated by the ultracentrifugation method, and their size and charge were measured with a TEM and zeta sizer. Then doxorubicin (Dox) loading into exosomes (PLT-Exo-Dox) was done by electroporation and incubated with MDA-MB-231 cells. In exosome characterization, CD62 positivity was higher in platelet pellets, while CD9 positivity was higher in released exosomes. The size of PLT-Exo and PLT-Exo-Dox was 82.02 ± 5.21 nm and 116 ± 3.73 nm, with a polydispersity index of 0.26 ± 0.04 and 0.39 ± 0.06, and the Zeta potential was − 16.45 mV and 24.07 mV, respectively. The encapsulation efficiency of the preparation was 86.02 ± 6.16