Serap Evran

In this study, development and optimization of a multiplex PCR assay for the simultaneous amplification of selected bacterial and fungal targets were performed. Source of hospital‐acquired infections‐specific, ( Staphylococcus aureus , Klebsiella pneumoniae , Pseudomonas aeruginosa , Candida albicans , Fusarium oxysporum , and Scedosporium prolificans ) primer sets targeting conserved genomic regions were systematically evaluated for potential nonspecific interactions, cross‐reactivity, and structural stability using in silico tools and empirical analyses. Multiplex PCR conditions were optimized using genomic DNA from reference strains under controlled laboratory conditions, with particular emphasis on annealing temperature, MgCl ₂ concentration, and primer balance. In addition, the effect of gold nanoparticles (AuNPs) on amplification performance was investigated. The inclusion of 1.0 nM 10 nm AuNPs enhanced overall amplification efficiency, with a more pronounced improvement observed for fungal targets. Overall, this study establishes an analytical proof of concept for a nanoparticle‐assisted multiplex PCR configuration capable of simultaneously amplifying multiple bacterial and fungal targets in a single reaction.

The plague, caused by Yersinia pestis, has resulted in significant mortality over the past century. Despite advances in antimicrobial therapy, plague remains a re-emerging infectious disease with ongoing outbreaks and increasing concerns regarding antimicrobial resistance. Today, plague cases are still being reported, and the loss of effectiveness of treatment methods remains a major challenge. Therefore, effective treatment strategies are needed. In this study, we aimed to develop aptamers specific to Yersinia outer protein M (YopM), a key immunosuppressive protein that is essential for virulence. Our goal was to develop an aptamer that binds to YopM and inhibits its interaction with the human DEAD-box helicase 3 (DDX3) protein. YopM-DDX3 protein interaction was targeted because of its key role in nucleocytoplasmic shuttling of YopM. To achieve this, we developed the YopM16 aptamer using magnetic bead-based (Systematic Evolution of Ligands by Exponential Enrichment) (SELEX). The selected YopM16 aptamer exhibited a half-maximal inhibitory concentration(IC₅₀) value of 103.3 ± 2 nM and effectively inhibited the interaction between YopM and DDX3. The inhibitory effect of the aptamer on protein interaction was confirmed using a pull-down assay and colorimetric test. Given that protein–protein interaction surfaces are considered undruggable, YopM16 is a promising inhibitor with the potential to serve as a molecular tool to investigate the virulence mechanism of YopM, as well as a novel antibacterial agent upon validation of its inhibition in cellular models.

Drug abuse is a major public problem in the workplace, traffic, and forensic issues, which requires a standardized test device to monitor on-site drug use. For field testing, the most important requirements are portability, sensitivity, non-invasiveness, and quick results. Motivated by this problem, a point of care (POC) test based on lateral flow assay (LFA) was developed for the detection of cocaine (COC) and methamphetamine (MET) in saliva which has been selected as the matrix for this study due to its rapid and non-invasive collection process. In the design strategy of an LFA test, the use of gold nanoparticles (AuNPs) with strong optical properties has been combined with the advantages of selecting aptamers under in vitro conditions, making it a highly specific and stable recognition probe for the detection of small molecules in saliva. The developed aptamer-based LFA in a competitive format, was able to detect COC and MET in synthetic saliva at concentrations as low as 5.0 ng/mL. After analytical performance studies, the test system also detected COC and MET in real patient samples, which was verified by chromatographic methods.

Bacteriophages are currently considered one of the most promising alternatives to antibiotics under the ‘One Health’ approach due to their ability to effectively combat bacterial infections. This study aimed to characterize Vibrio species in hatchery water samples collected from an aquaculture farm and investigate the biocontrol potential of their bacteriophages. Vibrio spp. (n = 32) isolates confirmed by LNA probe-based qPCR were used as hosts. Three Vibrio phages were isolated. IKEM_vK exhibited a broad host range, infecting V. harveyi (n = 8), V. alginolyticus (n = 2), V. azureus (n = 1), and V. ordalii (n = 1). IKEM_v5 showed lytic activity against V. anguillarum (n = 4) and V. ordalii (n = 1), while IKEM_v14 was specific to V. scophtalmi (n = 4). The morphological appearance of phages and their lytic effects on the host were visualized using scanning electron microscopy (SEM). All three phages remained relatively stable within the pH range of 6–11 and up to 60 °C. The lytic activities and biofilm inhibition capabilities of these phages against planktonic Vibrio cells support their potential applications in controlling vibriosis in aquaculture systems.

Epigenetic changes play an important role in the pathophysiology of autoimmune diseases such as allergic asthma, multiple sclerosis, lung diseases, diabetes, cystic fibrosis, atherosclerosis, rheumatoid arthritis, and COVID-19. There are three main classes of epigenetic alterations: posttranslational modifications of histone proteins, control by non-coding RNA and DNA methylation. Since histone modifications can directly affect chromatin structure and accessibility, they can regulate gene expression levels. Abnormal expression and activity of histone deacetylases (HDACs) have been reported in immune mediated diseases. Increased acetylated levels of lysine residues have been suggested to be related to the overexpression of inflammatory genes. This review focuses on the effect of HDAC modifications on histone and non–histone proteins in autoimmune diseases. Furthermore, we discuss the potential therapeutic effect of HDAC inhibitors (HDACi) used in these diseases.

In recent years, magnetic nanoparticles (MNPs) have been widely used in many fields due to their advantageous properties, such as biocompatibility, easy modifiability, and high chemical stability. One of these areas is the detection of cancer. It is essential to use existing biomarkers, such as microRNAs (miRNAs), for the early diagnosis of this disease. miRNAs are challenging to distinguish and detect in biological samples because they are small, circulating molecules. It is necessary to use more sensitive and feature-rich systems. Thanks to their large surface areas and magnetic moments, MNPs allow easy separation of miRNA at low concentrations from complex samples (urine and blood) and rapid and specific detection in biosensing systems. Here, we discussed the synthesis and characterization methods of MNPs, their stabilization, and MNP-based biosensors in terms of miRNA detection. We considered the challenges and prospects of these biosensor systems in evaluating the development stages, sensitivity, and selectivity.

Diagnostic biomarkers based on epigenetic changes such as DNA methylation are promising tools for early cancer diagnosis. However, there are significant difficulties in directly and specifically detecting methylated DNA regions. Here, we report an electrochemical sensing system based on magnetic nanoparticles that enable a quantitative and selective analysis of the methylated septin9 (mSEPT9) gene, which is considered a diagnostic marker in early stage colorectal cancer (CRC). Methylation levels of SEPT9 in CRC samples were successfully followed by the selective recognition ability of a related peptide nucleic acid (PNA) after hybridization with DNA fragments in human patients’ serums and plasma (n = 10). Moreover, this system was also adapted into a point-of-care (POC) device for a one-step detection platform. The detection of mSEPT9 demonstrated a limit of detection (LOD) value of 0.37% and interference-free measurement in the presence of branched-chain amino acid transaminase 1 (BCAT1) and SRY box transcription factor 21 antisense divergent transcript 1 (SOX21-AS1). The currently proposed functional platform has substantial prospects in translational applications of early CRC detection.