Cenk Serhan Ozverel

Cat flea (Ctenocephalides felis) infestations continue to pose major health and economic burdens in veterinary settings. Fluralaner, an isoxazoline-class ectoparasiticide, effectively targets GABA-gated chloride (GABA-Cl) channels in fleas; however, concerns about hepatotoxicity and bioavailability limitations underscore the need for more effective, safe, and naturally derived alternatives. This study aimed to evaluate the antiparasitic potential and safety profiles of five flavonoids, quercetin, kaempferol, luteolin, apigenin, and naringenin, compared to Fluralaner through a comprehensive in silico approach. The interactions of flavonoids and Fluralaner with the C. felis GABA-Cl receptor were modeled using AutoDock Vina and validated via molecular dynamics simulations in GROMACS. Binding energies were calculated using the MM/PBSA method. Toxicological (LD₅₀, hepatotoxicity, carcinogenicity) and pharmacokinetic (Gastrointestinal (GI) absorption, P-glycoprotein substrate status) properties were predicted using SwissADME and ProTox-II. Naringenin and apigenin exhibited the strongest predicted binding affinities with stable ligand–receptor interactions. Among all compounds, naringenin demonstrated the most favorable binding energy, safety, and pharmacokinetic profile. While apigenin and kaempferol also showed promising characteristics, luteolin and quercetin were associated with moderate toxicity risks. Fluralaner presented comparable efficacy but raised concerns regarding hepatotoxicity and limited bioavailability. Naringenin would be a less toxic alternative to Fluralaner, exhibiting a better safety profile and greater efficacy.

Background/Objectives: Dental biofilms are complex microbial communities that could lead to the formation of dental caries and other oral diseases. This in vitro study aimed to evaluate the effects of two commercially available toothpastes containing bioactive glass (BioMin F and Sensodyne Repair & Protection) on biofilm removal from five contemporary restorative dental materials and bovine enamel. The objective of this study was to compare the effects of bioactive glass-containing toothpastes on Streptococcus mutans viability and colonization on different restorative materials and bovine enamel. Methods: A total of 42 disc specimens representing five restorative dental materials and bovine enamel were incubated with Streptococcus mutans for 24 h to develop biofilms. After bacterial adhesion, specimens were brushed with toothpaste slurries (1:2 dilution) and distilled water (control). Biofilm viability and bacterial colonization were evaluated using the MTT assay and colony-forming unit (CFU) counts, respectively. Biofilm morphology on selected specimens was examined by scanning electron microscopy (SEM). Differences among groups were analyzed statistically using Tukey’s test. Results: Statistically significant differences were observed among the study groups following brushing (p < 0.05). EQUIA Forte HT (EF) demonstrated the lowest levels of S. mutans viability and colonization, whereas ACTIVA PRONTO (AP) exhibited the highest levels. Among the toothbrushing methods, BioMin F resulted in a greater reduction in bacterial viability and colony counts independent of the restorative materials. Combined analysis indicated that EF brushed with Sensodyne Repair & Protect yielded the lowest bacterial burden, whereas AP brushed with distilled water exhibited the highest. The results describing bacterial viability and colony counts also correlate with the SEM images taken. Conclusions: Within the limitations of this in vitro study, Sensodyne Repair & Protect was associated with lower S. mutans viability and colonization on most restorative materials, particularly EQUIA Forte HT.

Since the development of immune checkpoint drugs targeting PD-1 and PD-L1, cancer immunotherapy has advanced significantly. However, there are drawbacks to using monoclonal antibodies for this purpose, such as their high manufacturing costs and potential immunological side effects. Small molecule inhibitors, such as Selective Estrogen Receptor Modulators (SERMs) could offer a potential alternative for cancer immunotherapy targeting PD-L1. This study aimed to evaluate the binding energies and interactions of 17 selected SERMs with the PD-L1 protein using molecular docking, Molecular Mechanics/Poisson-Boltzmann Surface Area (MM/PBSA) calculations, and umbrella sampling methods, in comparison to benchmark PD-L1 inhibitors. Raloxifene emerged as the top-performing compound, demonstrating superior binding affinities (-85.80 ± 1.20 kJ/mol, MM/PBSA; -84.90 ± 1.10 kJ/mol, umbrella sampling) compared to the BMS series of known PD-L1 inhibitors and other SERMs. Interaction analyses highlighted significant stabilizing interactions between raloxifene and key PD-L1 residues. Other SERMs, such as lasofoxifene and ormeloxifene, also demonstrated competitive binding affinities, reinforcing the versatility of SERMs as potential PD-L1 inhibitors. This preliminary study underscored the promise of SERMs as effective and accessible alternatives in cancer immunotherapy. However, further experimental validation is warranted to establish their efficacy in preclinical and clinical settings in the future.

The discovery of tumor‐associated antigens has been a significant milestone for cancer therapy as the current conventional therapies (radiotherapy, chemotherapy) were found to be insufficient in terms of cancer treatment. Human epidermal growth factor receptor (HER2) is a tumor‐associated antigen, aberrantly found in various cancers, including breast cancer. Today, most treatment strategies are skewed toward combination therapies rather than monotherapies, as they have been found primarily ineffective due to patients' resistance mechanisms. A combination of multiple therapeutic approaches (passive/active immunotherapy, conventional therapies) was found to be more effective in the clinic. This study investigated various vaccine combinations against the 4 T1‐HER2 xenograft model of Balb/c mice. Eight different groups of vaccine formulations were tested to find the best synergistic combination, including combinations of BM‐DC‐based vaccine, peptide‐based vaccine, anti‐PD‐L1, Doxorubicin, and QS‐21 adjuvant. Triple combination groups of immunotherapies exerted better activities in terms of tumor dimensions as Doxorubicin+BM‐DC‐HER2/Neu + QS‐21 + anti‐PD‐L1 and Doxorubicin+HER2/Neu + QS‐21 + anti‐PD‐L1 groups significantly decreased tumor dimensions. Similar groups also demonstrated a better stimulation of lymphocytes and IFN‐γ cytokine in the flow cytometry study, inducing a HER2/Neu‐specific antibody response in ELISA studies. Doxorubicin+BM‐DC‐HER2/Neu + QS‐21 + anti‐PD‐L1 combination demonstrated significantly higher specific cytotoxicity in lactate dehydrogenase data, as well as the CD4+ and CD8+ responses in the immunohistochemical study. In conclusion, these data indicated that the Doxorubicin+BM‐DC + HER2/Neu + QS‐21 + anti‐PD‐L1 vaccine combination synergistically generated a promising activity in terms of tumor dimension and immune response stimulation against the HER2 overexpressing breast cancer model in mice.

Olive oil, a cornerstone of the Mediterranean diet, is renowned for its high content of phenolic compounds and its protective effects on human health. However, variations in production methods and raw material quality can significantly influence its chemical composition and biological activity. Kara yağ, a traditionally produced, unrefined olive oil unique to Northern Cyprus, is obtained through ancient artisanal extraction techniques and is rich in bioactive phenolics. This study aimed to investigate and compare the effects of Kara yağ and extra virgin olive oil (EVOO) on proliferation, viability, and apoptosis in HCT-116 human colon cancer cells. Samples of Kara yağ and EVOO were obtained from local producers in Northern Cyprus. Their physicochemical parameters, including free acidity, peroxide value, moisture content, and water activity, were analyzed in accordance with the Turkish Food Codex standards. Total phenolic content was determined using the Folin–Ciocalteu method, and antioxidant capacity was assessed spectrophotometrically. The high peroxide value (121.35 meq/kg) of Kara yağ is very high, which means that it was heavily oxidized throughout its conventional manufacturing process, and that heat treatment enhanced the chemical breakdown. Ochratoxin A (2.6 µg/kg) and aflatoxin B1 (0.8 µg/kg) were found in low levels of Kara yağ and therefore, the study is one of the first in the literature to analyze the toxin profile of the Kara yağ. The sensory analysis revealed that EVOO had distinct positive flavor qualities, but Kara yağ did not have them and high concentration of defects. HCT-116 cells were treated with different concentrations (1:1, 1:2, and 1:4) of each oil for 24, 48, and 72 hours. Cell viability was assessed by MTT assay, while morphological and apoptotic changes were evaluated by microscopy and immunocytochemical staining for Ki-67, Caspase-3, and c-Myc. Kara yağ exhibited significantly higher total phenolic content (278.95 mg GAE/kg) and antioxidant activity compared to EVOO. Treatment with Kara yağ led to a marked reduction in cell viability and proliferation in a time- and dose-dependent manner. Enhanced apoptotic activity, characterized by increased caspase-3 expression and decreased Bcl-2 levels, was observed following Kara yağ exposure. Both oils showed comparable c-Myc staining patterns. The findings suggest that Kara yağ, due to its superior phenolic composition and antioxidant potential, exerts stronger cytotoxic and pro-apoptotic effects on HCT-116 colon cancer cells than extra virgin olive oil. These results highlight the potential of traditionally produced Kara yağ as a natural source of bioactive compounds with promising chemopreventive and therapeutic properties against colon cancer. Further studies are warranted to elucidate the underlying molecular mechanisms and confirm its safety and efficacy in vivo.

Root canal infections, particularly those involving Enterococcus faecalis and Candida albicans , continue to pose significant challenges in endodontic therapy due to both their inherent resistance to conventional agents. This study aimed to evaluate the antimicrobial efficacy of a newly synthesized piperazine–chalcone hybrid compound, both individually and in various formulations with calcium hydroxide, against these clinically relevant pathogens. The antimicrobial activities were assessed using the disk diffusion method over 24 and 48 h, whereas in silico analyses were conducted to further investigate the compound's molecular interactions, pharmacokinetics, and toxicity potential. Results demonstrated that the piperazine–chalcone hybrid exhibited strong antimicrobial activity against both pathogens, with inhibition zone diameters ranging from approximately 26–32 mm against E. faecalis and 18–28 mm against C. albicans , exceeding those observed for calcium hydroxide and standard antimicrobial controls. Molecular docking analysis revealed favorable binding interactions of the hybrid compound with d ‐alanine: d ‐alanine ligase, with a docking score of −8.6 kcal/mol. In silico toxicological assessment predicted a favorable safety profile, with an LD ₅₀ value >5000 mg/kg and no major toxicity alerts. These findings suggest that the piperazine–chalcone hybrid holds promise as a safe and effective candidate for enhanced endodontic disinfection.

Antibiotic resistance poses a significant global health threat, necessitating innovative strategies to combat bacterial infections. β-lactamases, enzymes that hydrolyze β-lactam antibiotics, contribute significantly to antibiotic resistance. Phage therapy, utilizing bacteriophages to target β-lactamases, holds promise as an alternative treatment approach. In this study, we investigated the binding affinities between extended-spectrum β-lactamases (ESBLs), metallo-β-lactamases (MBLs), and AmpC-type β-lactamases with marine phages. Molecular modeling, molecular docking, and molecular dynamics simulations were employed using I-TASSER, AutoDock Vina, and GROMACS software to explore the interactions between tail tube proteins of selected marine phages (P-SSM2, P-SSM4, and P-SM2) and various β-lactamases. Among ESBLs, KPC-2 consistently exhibited the highest binding free energy, indicating robust interactions with marine phages. Similarly, among all MBLs, NDM-1 and VIM-1 showed superior binding free energies across all phage complexes. In addition, among AmpC-type beta-lactamases, CMY-2 displayed higher binding affinities across various phage complexes than DHA-1. In silico analyses suggest that marine phage tail proteins may interact with β-lactamases in a manner that could potentially mitigate antibiotic resistance. The differential binding affinities observed highlight the versatility of marine phages in targeting specific beta-lactamases. This novel approach could lead to the development of novel therapeutic strategies for multidrug-resistant gram-negative infections.

West Nile Virus (WNV) infection represents a major global public health challenge. Even though most of the patients are asymptomatic, some cases progress to critical condition which may be fatal. Diagnosis traditionally relies on serological methods, but their limitations, including cross‐reactivity, highlight the need for alternative approaches. Here, we present the development and validation of a novel RT‐qPCR assay for precise and rapid detection of WNV RNA in urine, emerging as a promising specimen due to its noninvasive collection and high viral load. The assay demonstrates high efficiency and sensitivity, with a detection limit comparable to commercially available kits. This study highlights the importance of in‐house kit design as a diagnostic tool in regions affected by emerging tropical infections, such as WNV, exemplified Cyprus. It emphasizes the critical role of low‐cost, early detection with high sensitivity and specificity in infection control and surveillance efforts.

Objective: The primary goal of this study was to assess the binding characteristics and affinities of melatonin and certain U.S. Food and Drug Administration (FDA)-approved lipophilic drugs used in the treatment of multiple sclerosis (MS) with the brain and muscle Arnt-like 1 (BMAL1) clock protein. Additionally, the study aimed to investigate their potential as modulators of BMAL1 activity for therapeutic applications in MS.Methods: Molecular docking and simulation studies were conducted to investigate the interactions between the BMAL1 protein and the selected agents as ligands. Docking simulations were performed using AutoDock Vina, and key interactions were analyzed with Biovia Discovery Studio Visualizer. A molecular dynamics simulation was conducted using GROMACS 2020.4 and the binding free energies were calculated using the molecular mechanics/Poisson-Boltzmann surface area (MM/PBSA) method. The pharmacokinetic properties of the ligands were predicted using the BOILED-Egg model implemented in the SwissADME web tool.Results: Melatonin demonstrated the highest binding affinity (-8.5 kcal/mol) and most favorable binding free energy (-245.26±1.27 kJ/mol) with BMAL1, suggesting its potential synergistic activity with other FDA-approved drugs in MS therapy. The pharmacokinetic analysis indicated that all three ligands were likely to cross the blood-brain barrier and exhibit high gastrointestinal absorption, with teriflunomide and melatonin potentially achieving a better profile.Conclusion: This study highlights the potential of concomitant use of melatonin to modulate BMAL1 activity, providing a foundation for developing targeted chronotherapeutic strategies in MS treatment.