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In this paper, three different Zn(II) complexes with (E)-2-(2-(1-(6-bromopyridin-2-yl)ethylidene)hydrazinyl)-N,N,N-trimethyl-2-oxoethan-1-aminium chloride (HLCl) have been synthesized and characterized by single crystal X-ray diffraction, elemental analysis, IR and NMR spectroscopy. All complexes are mononuclear, with the ligand (L) coordinated in a deprotonated formally neutral zwitterionic form via NNO donor set atoms. Complex 1 forms an octahedral geometry with the composition [ZnL2](BF4)2, while complexes 2 [ZnL(NCO)2] and 3 [ZnL(N3)2] form penta-coordinated geometry. Density functional theory (DFT) calculations were performed to enhance our understanding of the structures of the synthesized complexes and the cytotoxic activity of the complexes was tested against five human cancer cell lines (HeLa, A549, MDA-MB-231, K562, LS 174T) and normal human fibroblasts MRC-5. Additionally, antimicrobial and antifungal activity of these complexes was tested against a panel of Gram-negative and Gram-positive bacteria, two fungal strains, and a yeast strain. It is noteworthy that all three complexes show selective antifungal activity comparable to that of amphotericin B. Molecular docking analysis predicted that geranylgeranyl pyrophosphate synthase, an enzyme essential for sterol biosynthesis is the most likely target for inhibition by the tested complexes.

Acute myeloid leukemia (LAML) is one of the most prevalent hematological malignancies. In recent years, while targeted approaches have shown promise in the fight against cancer, the treatability and prognosis of patients remain inadequate due to the shortage of drugs. Noncoding RNAs, especially circular RNA (circRNA) and microRNA (miRNA), have been shown to play a unique role in tumor development. This study aims to identify the disease-associated circRNA–miRNA– mRNA network by bioinformatic analysis and investigate the mechanisms in the development and progression of LAML. Additionally, it reveals the promising roles of these molecules as a diagnostic biomarker and therapeutic target for LAML treatment. Using various bioinformatics approaches, we identified the hsa_circ_0058058/miR-324-5p axis in LAML and its possible functions in LAML development. According to our results, hsa circ-0058058 can regulate the expression of AP1G1 and SP1 through miR-324-5p to support angiogenesis, the cell cycle, and DNA replication processes. Downregulation of hsa circ-0058058 may contribute to the anticancer functions of miR-324-5p on LAML tumorigenesis, and upregulation of miR-324-5p can abolish the oncogenic effects of AP1G1 and SP1 on LAML tumorigenesis. Additionally, highly enriched pathways indicated possible interactions between molecules underlying LAML pathology. Targeted molecules within this network may be able to function as therapeutic and diagnostic biomarkers for disease, while more research and clinical confirmation are needed.

Radiation‑induced normal tissue toxicity is a common acute and chronic outcome of radiotherapy (RT) for prostate cancer (PCa). There are currently no existing pre‑assessments before treatment to predict acute and late RT‑induced toxicity. Novel predictive blood biomarkers in radiation oncology may improve treatment decision‑making and therapeutic monitoring for patients with PCa. A comprehensive systematic search of microRNA (miRNA/miR) profiling studies published in PubMed, Science Direct and Google Scholar was performed. The present systematic review, supported by meta‑analysis, summarises key findings and discusses the results of prospective clinical studies investigating miRNA expression levels and their association with RT‑induced toxicity in PCa. Out of seven clinical studies, six highlighted levels of 10 miRNAs changing in plasma, serum or peripheral blood mononuclear cells during RT. The post‑RT expression levels of miRNA‑132‑5p, miRNA‑1‑3p, miRNA‑410 and miRNA‑221 were increased, and miRNA‑23a‑3p expression was decreased among patients with low‑grade RT‑induced toxicity. Furthermore, in patients with high‑grade RT toxicity, miRNA‑197‑3p, miRNA‑151a‑5p, miRNA‑18b‑5p, miRNA‑99a and miRNA‑21 expression was increased, while miRNA‑132‑5p expression was decreased. The present miRNA meta‑analysis could be the focus of future studies to identify their potential clinical value as PCa biomarkers and therapeutic mediators in RT‑induced toxicity. The variations in miRNA levels post‑RT could be used as predictive biomarkers of RT‑induced toxicity. However, further extensive validation is required to establish the relationship between miRNA expression levels and RT‑induced toxicity in PCa and to confirm their predictive value.

Variations in the hepatitis C virus (HCV) core gene are related to the progression of liver fibrosis and therapy response. However, the influence of individual amino acid (aa) substitutions at different positions of HCV Core protein on the response to combined pegylated interferon/ribavirin (PEG-IFN/RBV) therapy and disease progression is not yet fully understood. The HCV Core protein can inactivate various genes in the host genome by affecting the methylation of their promoters, leading to liver damage and carcinogenesis. Two genes whose methylation status is affected by the HCV Core protein are the tumor suppressor genes RASSF1A and p16. We have previously shown that the methylation status of these two genes, together with the single nucleotide polymorphism (SNP) rs12979860 near the interleukin-28 beta subunit (IL-28B) gene, influences the response to combined therapy. Herein, we investigated a possible association between detected aa substitutions in HCV Core protein and response to combined therapy, liver disease progression, IL28B genotype, and the methylation status of the RASSF1A and p16 genes. In 29 examined patients we found no association between individual aa substitutions and therapy response. However, we observed that patients with HCV Core aa substitutions at position 75 and CT/TT IL28B genotypes were non-responders (NR) (p=0.023), which was associated with the presence of unmethylated RASSF1A gene. In contrast, even 75% of patients with aa substitutions at position 91 and CC IL28B genotype achieved a sustained virologic response (SVR) (p=0.030), and 70% of them had methylated RASSF1A gene. There was no significant association between the methylation status of the p16 gene and aa variations in the HCV core region. Our results suggest that combined analysis of aa substitutions in HCV Core protein, IL28B genotype, and methylation status of the RASSF1A gene may help predict response to PEG-IFN/RBV therapy in patients with chronic hepatitis C genotype 1b.x

Variations in the hepatitis C virus (HCV) core sequence have been related to disease progression and response to antiviral therapy. Previously we showed that the methylation status of RASSF1A and p16 genes, and IL28B genotypes affects the response to pegylated interferon/ribavirin (PEG-IFN/RBV) therapy. Herein we investigated whether amino acid (aa) substitutions in the HCV core region alone or in combination with IL28B genotypes and RASSF1A/p16 methylation affect the response to PEG-IFN/RBV therapy and liver disease progression. Among 29 examined patients, we found no association between single aa substitutions and response to therapy. However, we observed that patients with the HCV core aa substitution at position 75 and CT/TT IL28B genotypes were non-responders (NR), (P=0.023). Moreover, these patients had unmethylated RASSF1A. In contrast, most patients (75%) with aa substitutions at position 91 and CC IL28B genotype achieved sustained virologic response (SVR), (P=0.030), and 70% of them had methylated RASSF1A gene. Our results suggest that combined analysis of aa substitutions in the core protein, the IL28B rs12979860 polymorphism, and the methylation status of the RASSF1A gene may help in predicting treatment response to PEG-IFN/RBV in genotype 1b chronic hepatitis C patients.

Background: It has been demonstrated that aromaƟ c plant essenƟ al oils (EO) contain phytochemicals with anƟ - infl ammatory, anƟ oxidant, and anƟ cancer acƟ viƟ es. The goal of this study was to examine the cytotoxicity and the anƟ cancer acƟ on mechanisms of an EO extracted from Satureja montana plant species, originaƟ ng from Montenegro. Material and methods: The cytotoxic acƟ vity was assessed against human cancer cell lines: cervical adenocarcinoma HeLa, malignant melanoma A375, colorectal adenocarcinoma LS 174T, lung carcinoma A549, as well as against normal human lung fi broblasts MRC-5 by MTT assay. Using fl ow cytometry, it was examined how HeLa cells were distributed throughout the cell cycle aŌ er treatment with EO and whether caspase-3, caspase-8, and caspase-9 were acƟ vated. RT-qPCR was used to assess the expression levels of genes and miRNA in HeLa cells. Results: Satureja montana EO exerted strong cytotoxicity against human cancer cell lines, with IC50 values ranging from 0,12 to 0,18 µL/mL, The strongest cytotoxic acƟ vity of EO was observed against lung carcinoma A549 cells with an IC50 value of 0.12 µL/mL and LS 174T colorectal adenocarcinoma cells with IC50 value of 0.13 µL/mL. The lowest cytotoxicity was determined against normal lung fi broblasts MRC-5 (IC50 value of 0.19 µL/mL). AŌ er 24 hours of treatment with Satureja montana EO, a striking increase in the proporƟ on of HeLa cells in the subG1 phase was seen in comparison with control cells. Pretreatment of HeLa cells with caspase inhibitors showed that Satureja montana EO acƟ vated all three invesƟ gated caspases to trigger apoptosis. In comparison to control cells, Satureja montana EO treatment of HeLa cells decreased MMP2 gene expression levels, and elevated MMP9 and VEGFA gene expression levels. Satureja montana EO treatment increased the expression levels of the tumor supressive miR-16 and miR-34ain HeLa cells, and increased levels of miR21, with oncogenic role in cervical cancer. Concl

Introduction: The incidence of lung cancer (LC) in Serbia has increased over the last three decades. Up to 40% of patients with advanced non-small cell lung cancer (NSCLC) develop pleural effusion (PE). The aim of this study was to evaluate the usability of existing methods for isolation of extracellular vesicles from PE samples of patients with advanced non-small cell lung cancer and to characterize them for further use in the clinical/diagnostic/research setting. Material & Methods: PE samples diluted in PBS (1:1) from patients with advanced NSCLC were used. In-house spherical porous methacrylate-based copolymer coupled with VHH antibodies was used for the isolation of extracellular vesicles (EV) from PE samples. Flow-cytometry was performed for detection of exosomal markers. Results: A pool of PE was prepared from 5 patients with advanced NSCLC. Flow cytometry confirmed that the isolation of EVs was successful using the in-house affinity chromatography method. The presence of CD9 antigen was detected, as well as a decrease in the signal after the addition of Triton X-100. Further plans include the analysis of CD63 and CD81 antigens using flow cytometry, NTA analysis to determine the number and diameter of obtained vesicles, as well as TEM and SEM to determine their morphology. Discussion: We aim to investigate for the first time whether this method is applicable to pleural effusion samples as a new cancer liquid biopsy sample type. We also plan to evaluate the method in comparison with a commercial isolation kit.

Introduction Nearly 60% of patients with breast cancer (BC) undergo radiotherapy (RT). A significant proportion of patients experience side effects that can influence their quality of life and treatment effectiveness. Gene and microRNA expression patterns may be associated with individual patients' sensitivity to RT. Material and Methods Expression levels of miR-133b, miR-206, and their target genes matrix metalloproteinase 9 (MMP9) and vascular endothelial growth factor (VEGFA) were investigated in normal tissue adjacent to tumors of 27 BC patients. Sixteen out of 27 (59.3%) patients underwent RT, while 13 of them had breast skin reactions (acute dermatitis). Patients with acute dermatitis were divided into two groups, H-hypofractionated and F-standardly fractionated group. The H group underwent 16 fractions of RT (2.65 Gy), while the second group-F had 25 fractions (2 Gy) or 25 fractions plus boost. All patients from the F group had acute dermatitis before the 25th fraction, so the boost did not influence the time of toxicity occurrence. After the RNA extraction, followed by reverse transcription, expression levels of the four molecules were measured by RT-qPCR. Relative quantity (RQ) units were calculated by a comparative 2-ddCt method. Results and Discussions MiR-133b/206 and MMP9/VEGFA gene levels did not differ between patients from H and F groups (p = 0.573; p = 0.143; p = 0.727; p = 0.582, respectively). The F group was divided into two subgroups-earlier (E) subgroup with patients with skin reactions before or at 20th fraction, and the later (L) subgroup with patients with skin reactions after 20th fraction. VEGFA gene expression was significantly higher in the E group compared with the L group, indicating that it may be a specific predictor of earlier onset of acute skin reaction in the normal breast tissue of patients undergoing RT (p = 0.036, Mann–Whitney U test). MicroRNAs miR-133b, miR-206, and MMP9 were not associated with earlier or later onset of ski

Introduction The phytochemicals present in essential oils derived from aromatic plants of genus Thymus have been reported to exert antioxidant, anti-inflammatory, and anticancer effects. The aim of the research was to examine the cytotoxic activity and the mechanisms of anticancer action of the two essential oils obtained from Thymus vulgaris and Thymus serpyllum grown in Montenegro. Material and Methods The cytotoxic activity was determined against four human cancer cell lines: cervical adenocarcinoma HeLa, malignant melanoma A375, colorectal adenocarcinoma LS 174T, and lung carcinoma A549, as well as against normal lung fibroblasts MRC-5 by MTT assay. The cell cycle phase distribution of HeLa cells and the potential activation of caspase-3, caspase-8, and caspase-9 were investigated by flow cytometry. Gene and microRNA expression levels in HeLa cells were measured using RT-qPCR. The intracellular levels of reactive oxygen species (ROS) in MRC-5 cells were measured by flow cytometry. Results and Discussions Both essential oils exerted strong cytotoxicity against cancer cell lines with IC50 concentrations in the range from 0,20 to 0,24 µL/mL for T. vulgaris and from 0.32 to 0.49 µL/mL for T. serpyllum. Strong cytotoxicity was observed against lung fibroblasts MRC-5. The remarkable increases in the percentage of HeLa cells in the subG1 phase of the cell cycle after 24 h treatment with T. vulgaris and T. serpyllum essential oils were observed in comparison to the control cells. Pretreatment of HeLa cells with caspase inhibitors showed that T. vulgaris oil induced apoptotic cell death through caspase-3 and caspase-8, while T. serpyllum oil induced apoptosis through caspase-3 activation. Both essential oils decreased intracellular ROS levels in MRC-5 cells and reduced levels of oxidative stress induced by hydrogen peroxide. The treatment of HeLa cells with T. vulgaris oil lowered the MMP2 expression levels, increased MMP9 and VEGFA levels when compared with control cell

Introduction The plant species belonging to the large genus Helichrysum are a valuable source of bioactive compounds. The aim of this study was to investigate the anticancer properties of the two Helichrysum italicum extracts obtained from plant material by supercritical CO2 extraction. Material and Methods The cytotoxicity of the extracts was examined against six human cancer cell lines: cervical adenocarcinoma HeLa, lung carcinoma A549, prostate adenocarcinoma PC-3, breast adenocarcinoma MCF-7, melanoma A375, and chronic myelogenous leukemia K562, as well as against two human normal cell lines: lung fibroblasts MRC-5 and keratinocytes HaCaT. Cell cycle analysis was performed by flow cytometry. Gene and microRNA expression levels were measured by RT-qPCR. Results and Discussions The H. italicum extracts exerted concentration-dependent and selective cytotoxic effects on cancer cells. HeLa, A375, and K562 cells were the most sensitive to the cytotoxic activity of both extracts. The extracts showed lower intensities of cytotoxic activity against normal HaCaT cells when compared with intensities of activity against K562, A375, and HeLa cells. An increases in the percentages of cells within subG1 and S phases of the cell cycle were observed in HeLa cells incubated for 24 h with IC50 and 2IC50 concentrations of the extracts in comparison with control cells. A similar effect on the cell cycle phase distribution was seen in A375 cells after 24 h incubation. The G2/M phase arrest was also detected in A375 cells exposed to IC50 concentrations of the extracts. Both extracts induced apoptosis in HeLa cells through the activation of effector caspase-3. The extracts triggered apoptosis through the intrinsic pathway mediated by caspase-9 and the extrinsic pathway mediated by caspase-8. Each of the extracts decreased expression levels of MMP2 in HeLa cells, slightly increased levels of MMP9, and increased levels of VEGFA. Up-regulations of genes involved in cancer invasion and pro

COVID-19 is a viral respiratory infection induced by the newly discovered coronavirus SARS-CoV-2. miRNA is an example of a strong and direct regulator of a gene’s transcriptional activity. The interaction between miRNAs and their target molecules is responsible for homeostasis. Virus-derived and host-derived miRNAs are involved in the activity of hiding from immune system cells, inducing the inflammatory reaction through interplay with associated genes, during SARS-COV-2 infection. Interest in miRNAs has raised the comprehension of the machinery and pathophysiology of SARS-COV-2 infection. In this review, the effects and biological roles of miRNAs on SARS-CoV-2 pathogenicity and life cycle are described. The therapeutic potential of miRNAs against SARS-CoV-2 infection are also mentioned.

Type 2 diabetes mellitus (T2DM) is associated with functional deterioration of the salivary gland and dental pulp, related to oxidative stress. The aim was to integrate experimental and bioinformatic findings to analyze the cellular mechanism of melatonin (MEL) action in the human parotid gland and dental pulp in diabetes. Human parotid gland tissue was obtained from 16 non-diabetic and 16 diabetic participants, as well as human dental pulp from 15 non-diabetic and 15 diabetic participants. In human non-diabetic and diabetic parotid gland cells (hPGCs) as well as in dental pulp cells (hDPCs), cultured in hyper- and normoglycemic conditions, glial cell linederived neurotrophic factor (GDNF), MEL, inducible nitric oxide synthase (iNOS) protein expression, and superoxide dismutase (SOD) activity were measured by enzyme-linked immunosorbent assay (ELISA) and spectrophotometrically. Bioinformatic analysis was performed using ShinyGO (v.0.75) application. Diabetic participants had increased GDNF and decreased MEL in parotid (p < 0.01) and dental pulp (p < 0.05) tissues, associated with increased iNOS and SOD activity. Normoglycemic hDPCs and non-diabetic hPGCs treated with 0.1 mM MEL had increased GDNF (p < 0.05), while hyperglycemic hDPCs treated with 1 mM MEL showed a decrease in up-regulated GDNF (p < 0.05). Enrichment analyses showed interference with stress and ATF/CREB signaling. MEL induced the stress-protective mechanism in hyperglycemic hDPCs and diabetic hPGCs, suggesting MEL could be beneficial for diabetes-associated disturbances in oral tissues.

Prevention of chronic hepatitis C (CHC) and its complications is based on antiviral therapy and early detection of reliable molecular markers in persons under risk. We investigated whether the methylation status of RASSF1A and p16 genes, alone or in combination with host and viral factors, affects the response to therapy with pegylated interferon/ribavirin (PEG-IFN/RBV). Methylation-specific polymerase chain reaction (MSP) was used to determine the methylation status of the target promoter sequences of RASSF1A and p16 in circulating-free DNA from the peripheral blood of 49 patients with CHC genotype 1b. The methylation status of the examined genes did not affect the response to therapy. However, the simultaneous presence of either RASSF1A or p16 methylation and the CC genotype of IL28B was significantly related to a sustained virologic response (P=0.009 and P=0.032, respectively). After Bonferroni correction, only the result concerning the RASSF1A gene remained significant (P<0.0125). Methylation of RASSF1A was associated with the CC genotype of the IL28B gene (P=0.024) and a higher viral load (≥400 000 IU/mL, P=0.009). Our results suggest that combined analysis of RASSF1A gene methylation and IL28B rs12979860 polymorphism could potentially help in the prediction of therapy response in CHC genotype 1b patients.

Non-coding RNAs (ncRNAs) represent a research hotspot by playing a key role in epigenetic and transcriptional regulation of diverse biological functions and due to their involvement in different diseases, including oral inflammatory diseases. Based on ncRNAs’ suitability for salivary biomarkers and their involvement in neuropathic pain and tissue regeneration signaling pathways, the present narrative review aims to highlight the potential clinical applications of ncRNAs in oral inflammatory diseases, with an emphasis on salivary diagnostics, regenerative dentistry, and precision medicine for neuropathic orofacial pain.

Phytochemicals and bioactive substances derived from a wide range of plant extracts have been reported to exert various anticancer effects. Prostate cancer is one of the leading causes of cancer-related deaths within the male population. Prostate cancer-specific miRNA signatures were associated with cancer formation and progression, with various subtypes, and response to therapy. MicroRNA levels of expression were shown to change after the treatment of various compounds and substances extracted from natural products. Natural herbal compounds were shown to induce variations in miRNA expression levels in cancer cells. The aims of this study were to investigate the cytotoxic effects of methanol, ethyl-acetate, and hexane extracts obtained from branch-body part and flowers of Hypericum perforatum L. against humane PC-3 and DU 145 and to test potential miRNA-128/133b/155/193a/206/21/335 signature changes and differences between the two prostate cancer cell lines. Cytotoxic activity of H. perforatum extracts, their effects on cell cycle distribution, and miRNA expression levels were examined in humane PC-3 and DU 145 prostate cancer cells by MTT cell survival assay, flow cytometry, and quantitative real-time PCR. Hexane extract of flowers showed the strongest intensity of cytotoxic activity against PC-3 and DU 145 cells. The highest increase in the percentage of PC-3 cells in the subG1 phase was observed in cell samples treated with hexane extract of flowers and branch-body part. Significant differences in miRNA-128/133b/155/193a/206/21/335 levels were observed between PC-3 and DU 145 cell lines, especially in samples treated with flower extracts compared with the branch-body part. Conclusions: Investigated extracts have significant anticancer potential not only from the aspects of cytotoxicity and cell cycle effects but also from the aspect of lowering oncogenic or increasing tumor-suppressive miRNAs. The best effect might be the increase of tumor-suppressive miR-128 (ac

Breast cancer (BC) is the most common heterogeneous disease in women and one of the leading causes of cancer-related death. Surgery, chemotherapy, radiotherapy, hormone, and targeted therapy are the gold standards for BC treatment. One of the significant challenges during the treatment of BC represents resistance to chemotherapeutics, resistance that severely limits the use and effectiveness of the drugs used for BC treatment. Therefore, it is essential to develop new strategies to improve therapeutic efficacy. Circular RNAs (circRNAs) are a large group of non-coding RNAs that covalently form closed circular loops by joining their 5′, and 3′; ends. Accumulating evidence suggests that circRNAs have a vital role in cancer development, progression, and BC resistance to chemotherapy. The purpose of this review is to discuss the biological properties of circRNAs, and how circRNAs induce resistance to conventional therapeutic anti-cancer drugs used in BC treatment, by emphasizing and summarizing the potential roles of circRNAs in mechanisms of drug resistance, such as drug efflux, apoptosis dysfunction, autophagy, and DNA damage repair. CircRNAs are associated with drug resistance via ATP-binding cassette (ABC) efflux transporters, while some others by inhibition of cell apoptosis, thus leading to resistance to tamoxifen in BC cells. In contrast, others are involved in the promotion of BC cells chemoresistance by doxorubicin-induced autophagy. CircRNAs may have clinical significance in regulating or overcoming BC drug resistance and may give directions towards a novel approach to personalized BC treatment. CircRNAs may significantly contribute to the identification of new therapeutic targets for the prevention of BC chemoresistance. © 2022, Tech Science Press. All rights reserved.