Metallothioneins (MTs) are small cysteine-rich intracellular proteins. The best-known biological functions of MTs are sequestration of metal ions and maintenance of redox homeostasis. Despite these protective functions, it has been demonstrated that MTs are involved in tumorigenesis, cellular differentiation, drug resistance, and metabolic disorders such as diabetes and obesity, in which MTs expression is substantially deregulated in adipose tissue. In addition, many studies have experimentally evidenced a possible role of MTs in the development of diabetes. Given the rich biochemical properties of MTs, it can be concluded that they are involved in several aspects of development and progression of obesity and diabetes. Thus, evaluation of expression of MTs could serve as biomarker to personalize available therapeutic interventions and possibly to develop novel advanced therapeutic modalities. Overall, the purpose of this review is analyze and review the latest studies aimed on the multiple roles of MTs in metabolic disorders, possible use of MTs as obesity and diabetes biomarkers and the role of MTs in cardioprotection during diabetes progression.

• Publication type
• Journal Article MeSH
• Review MeSH

Alexander disease (AxD) is a rare and severe neurodegenerative disorder caused by mutations in glial fibrillary acidic protein (GFAP). While the exact disease mechanism remains unknown, previous studies suggest that mutant GFAP influences many cellular processes, including cytoskeleton stability, mechanosensing, metabolism, and proteasome function. While most studies have primarily focused on GFAP-expressing astrocytes, GFAP is also expressed by radial glia and neural progenitor cells, prompting questions about the impact of GFAP mutations on central nervous system (CNS) development. In this study, we observed impaired differentiation of astrocytes and neurons in co-cultures of astrocytes and neurons, as well as in neural organoids, both generated from AxD patient-derived induced pluripotent stem (iPS) cells with a GFAPR239C mutation. Leveraging single-cell RNA sequencing (scRNA-seq), we identified distinct cell populations and transcriptomic differences between the mutant GFAP cultures and a corrected isogenic control. These findings were supported by results obtained with immunocytochemistry and proteomics. In co-cultures, the GFAPR239C mutation resulted in an increased abundance of immature cells, while in unguided neural organoids and cortical organoids, we observed altered lineage commitment and reduced abundance of astrocytes. Gene expression analysis revealed increased stress susceptibility, cytoskeletal abnormalities, and altered extracellular matrix and cell-cell communication patterns in the AxD cultures, which also exhibited higher cell death after stress. Overall, our results point to altered cell differentiation in AxD patient-derived iPS-cell models, opening new avenues for AxD research.

• Keywords
• Alexander disease, GFAP, iPS cells, neural organoids,
• MeSH
• Alexander Disease * genetics   pathology   metabolism   MeSH
• Astrocytes * metabolism   pathology   MeSH
• Cell Differentiation * physiology   MeSH
• Glial Fibrillary Acidic Protein * metabolism   genetics   MeSH
• Induced Pluripotent Stem Cells * metabolism   MeSH
• Coculture Techniques MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Mutation MeSH
• Neural Stem Cells metabolism   MeSH
• Neurons metabolism   pathology   MeSH
• Organoids metabolism   pathology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• GFAP protein, human MeSH Browser
• Glial Fibrillary Acidic Protein * MeSH

Objectives: To advance our knowledge of disease mechanisms and therapeutic options, understanding cell cycle regulation is critical. Recent research has highlighted the importance of reactive oxygen species (ROS) in cell cycle regulation. Although excessive ROS levels can lead to age-related pathologies, ROS also play an essential role in normal cellular functions. Many cell cycle regulatory proteins are affected by their redox status, but the precise mechanisms and conditions under which ROS promote or inhibit cell proliferation are not fully understood.Methods: This review presents data from the scientific literature and publicly available databases on changes in redox state during the cell cycle and their effects on key regulatory proteins.Results: We identified redox-sensitive targets within the cell cycle machinery and analysed different effects of ROS (type, concentration, duration of exposure) on cell cycle phases. For example, moderate levels of ROS can promote cell proliferation by activating signalling pathways involved in cell cycle progression, whereas excessive ROS levels can induce DNA damage and trigger cell cycle arrest or cell death.Discussion: Our findings encourage future research focused on identifying redox-sensitive targets in the cell cycle machinery, potentially leading to new treatments for diseases with dysregulated cell proliferation.

• Keywords
• Cell cycle, cell cycle signaling, oxidative stress, proliferation, reactive oxygen species, redox state, redox-sensitive targets,
• MeSH
• Cell Cycle * MeSH
• Humans MeSH
• Oxidation-Reduction * MeSH
• DNA Damage MeSH
• Cell Proliferation MeSH
• Reactive Oxygen Species * metabolism   MeSH
• Signal Transduction MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Reactive Oxygen Species * MeSH

Increased awareness of the impact of human activities on the environment has emerged in recent decades. One significant global environmental and human health issue is the development of materials that could potentially have negative effects. These materials can accumulate in the environment, infiltrate organisms, and move up the food chain, causing toxic effects at various levels. Therefore, it is crucial to assess materials comprising nano-scale particles due to the rapid expansion of nanotechnology. The aquatic environment, particularly vulnerable to waste pollution, demands attention. This review provides an overview of the behavior and fate of metallic nanoparticles (NPs) in the aquatic environment. It focuses on recent studies investigating the toxicity of different metallic NPs on aquatic organisms, with a specific emphasis on thiol-biomarkers of oxidative stress such as glutathione, thiol- and related-enzymes, and metallothionein. Additionally, the selection of suitable measurement methods for monitoring thiol-biomarkers in NPs' ecotoxicity assessments is discussed. The review also describes the analytical techniques employed for determining levels of oxidative stress biomarkers.

• Keywords
• Aquatic organism, Glutathione, Mass spectrometry, Metallothionein, Oxidative stress,
• MeSH
• Antioxidants * metabolism   MeSH
• Biomarkers * metabolism   MeSH
• Water Pollutants, Chemical * toxicity   analysis   MeSH
• Glutathione metabolism   MeSH
• Metal Nanoparticles * toxicity   chemistry   MeSH
• Humans MeSH
• Metallothionein metabolism   MeSH
• Environmental Monitoring methods   MeSH
• Oxidative Stress * drug effects   MeSH
• Peptides toxicity   MeSH
• Aquatic Organisms drug effects   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Antioxidants * MeSH
• Biomarkers * MeSH
• Water Pollutants, Chemical * MeSH
• Glutathione MeSH
• Metallothionein MeSH
• Peptides MeSH

Bisphenol-A (BPA) has become a great concern due to its toxic effects. The present study investigated the retrieval action of zinc (Zn) and folic acid (FA) supplementation against BPA-induced reproductive toxicities in male albino mice. A total of seventy-five 25-28 day-old mice were divided into five equal groups (group A-E, 15 mice in each group). The mice were given normal rations (control, group A) or administered with daily doses of BPA at 50 mg/kg body weight (b.w.) (group B-E). The mice from groups C, D and E were supplemented with Zn (10 mg/kg b.w.), FA (3 mg/kg b.w.) and both in the feed, respectively, daily for 12 weeks. Blood samples were collected, and the sera were separated for biochemical and hormonal analyses. The standard method was followed to test the sperm motility and sperm count. The testis samples were processed for a routine histopathological study using haematoxylin and eosin staining. The sperm counts, motility, and serum testosterone significantly declined in the BPA-exposed animals, but dramatically increased after the Zn and FA supplementation. There was significant degeneration of the seminiferous tubules in the testes of the BPA-exposed mice, which was recovered moderately by the Zn and FA supplementation. The study shows the retrieval action of zinc and folic acid in the restoration of normal reproductive function in bisphenol-A exposed male mice.

• Keywords
• BPA, biochemical, histopathology, hormone, reproduction,
• Publication type
• Journal Article MeSH

Doxorubicin (DOX) is one of the most frequently used anticancer drugs in breast cancer treatment. However, clinical applications of DOX are restricted, largely due to the fact that its action disturbs the pro/antioxidant balance in both cancerous and non-cancerous cells. The aim of this study was to investigate the influence of fullerene (C60) in cell treatment by DOX on the proliferation of human breast cancer cells (MCF-7), concentration of metallothionein (MT) and superoxide dismutase (SOD), and SOD activity in these cells. The use of C60 in complexes with DOX causes a change in the level of cell proliferation of about 5% more than when caused by DOX alone (from 60⁻65% to 70%). The use of C60 as a DOX nanotransporter reduced the MT level increase induced by DOX. C60 alone caused an increase of SOD1 concentration. On the other hand, it led to a decrease of SOD activity. C60 in complex with DOX caused a decrease of the DOX-induced SOD activity level. Exposure of MCF-7 cells to DOX-C60 complexes results in a decrease in viable cells and may become a new therapeutic approach to breast cancer. The effects of C60 in complexes with DOX on MCF-7 cells included a decreased enzymatic (SOD activity) and nonenzymatic (MT) antioxidant status, thus indicating their prooxidant role in MCF-7 cells.

• Keywords
• breast tumors, doxorubicin, drug delivery systems, fullerene, nanoparticles, metallothionein, superoxide dismutase,
• MeSH
• Doxorubicin pharmacology   MeSH
• Fullerenes chemistry   MeSH
• Humans MeSH
• Metallothionein metabolism   MeSH
• MCF-7 Cells MeSH
• Nanoparticles chemistry   MeSH
• Cell Proliferation drug effects   MeSH
• Antineoplastic Agents pharmacology   MeSH
• Superoxide Dismutase-1 metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Doxorubicin MeSH
• Fullerenes MeSH
• Metallothionein MeSH
• Antineoplastic Agents MeSH
• SOD1 protein, human MeSH Browser
• Superoxide Dismutase-1 MeSH

Human metallothionein-3 (hMT-3), also known as growth inhibitory factor, is predominantly expressed in the central nervous system. hMT-3 is presumed to participate in the processes of heavy metal detoxification, regulation of metabolism and protection against oxidative damage of free radicals in the central nervous system; thus, it could play important neuromodulatory and neuroprotective roles. However, the primary functions of hMT-3 and the mechanism underlying its multiple functions in neuroblastoma have not been elucidated so far. First, we confirmed relatively high expression of hMT-3 encoding mRNA in biopsies (n = 23) from high-risk neuroblastoma subjects. Therefore, we focused on investigation of the impact of hMT-3 up-regulation in N-Myc amplifying neuroblastoma cells. The differentially up-regulated genes involved in biological pathways related to cellular senescence and cell cycle were identified using electrochemical microarray with consequent bioinformatic processing. Further, as experimental verification of microarray data, the cytotoxicity of the cisplatin (CDDP) was examined in hMT-3 and mock cells by MTT and clonogenic assays. Overall, our data strongly suggest that up-regulation of hMT-3 positively correlates with the genes involved in oncogene-induced senescence (CDKN2B and ANAPC5) or apoptosis (CASP4). Moreover, we identified a significant increase in chemoresistance to cisplatin (CDDP) due to hMT-3 up-regulation (24IC50: 7.5 vs. 19.8 μg/ml), indicating its multipurpose biological significance.

• Keywords
• apoptosis, chemoresistance, cisplatin, metallothionein, oncogene-induced senescence,
• Publication type
• Journal Article MeSH

In this study, we describe the establishment of the human papillomavirus 18-positive, stage II, grade 1, T2N0M0 head and neck tumor primary cell line derived from oral squamous cell carcinoma of a non-smoking patient by using two different protocols. Furthermore, a preparation of subpopulations derived from this primary cell line according to the cluster of differentiation molecules CD44/CD90 status using magnetic bead-based separation and their characterization was performed. Impedance-based real-time cell analysis, enzyme-linked immunsorbant assay (ELISA), wound-healing assay, flow-cytometry, gene expression analysis, and MTT assay were used to characterize these four subpopulations (CD44+/CD90-, CD44-/CD90-, CD44+/CD90+, CD44-/CD90-). We optimised methodics for establishement of primary cell lines derived from oral squamous cell carcinoma tissue samples and subsequent separation of mesenchymal (CD90+) and epithelial (CD90-) types of tumorous cells. Primary cell line prepared by using trypsin proteolysis was more viable than the one prepared by using collagenase. According to our results, CD90 separation is a necessary step in preparation of permanent tumor-tissue derived cell lines. Based on the wound-healing assay, CD44+ cells exhibited stronger migratory capacity than CD44- subpopulations. CD44+ subpopulations had also significantly higher expression of BIRC5 and SOX2, lower expression of FLT1 and IL6, and higher levels of basal autophagy compared to CD44- subpopulations. Furthermore, co-cultivation experiments revealed that CD44-/CD90+ cells supported growth of epithelial tumor cells (CD44+/CD90-). On the contrary, factors released by CD44+/CD90+ type of cells seem to have rather inhibiting effect. The most cisplatin-resistant subpopulation with the shortest doubling time was CD44-/CD90+, but this subpopulation had a low migratory capacity.

• Keywords
• carcinoma, cell line, coculture techniques, head and neck neoplasms, tumor,
• Publication type
• Journal Article MeSH

Even with significant advances in operative skills and adjuvant therapies, the overall survival of patients suffering with head and neck squamous cancers (HNSCC) is unsatisfactory. Accordingly, no clinically useful prognostic biomarkers have been found yet for HNSCC. Many studies analysed the expression of potential markers in tumour tissues compared to adjacent tissues. Nevertheless, due to the sharing of the same microenvironment, adjacent tissues show molecular similarity to tumour tissues. Thus, gene expression patterns of 94 HNSCC tumorous tissues were compared with 31 adjacent tissues and with 10 tonsillectomy specimens of non-cancer individuals. The genes analysed at RNA level using quantitative RT-PCR and correlated with clinico-pathological conditions were as follows: EGF, EGFR, MKI67, BCL2, BAX, FOS, JUN, TP53, VEGF, FLT1, MMP2, MMP9, MT1A and MT2A. The elevated MT2A, BAX, EGF and JUN expression was associated with the influence of tumour cells on the rearrangement of healthy tissues, as well as a significant shift in the BAX/BCL2 ratio. Our investigation also indicated that adjacent tissues play an important role in cancerogenesis by releasing several tumour-supporting factors such as EGF. A gradual increase in the metallothionein expression, from the lowest one in tonsillectomy samples to the highest ones in tumour samples, suggests that MT expression might be tissue reaction to the presence of tumour cells. The results of this study confirmed the significance of metallothionein in tumori-genesis and gave evidences for its use as a potential HNSCC biomarker. Furthermore, this study highlighted the importance of histologically normal tumour-adjacent tissue in prediction of HNSCC progress.

• Keywords
• Biological markers, Gene expression, Head and neck neoplasms, Matrix metalloproteinase 9, Metallothionein, Prognosis, Tumour microenvironment,
• MeSH
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Biomarkers, Tumor biosynthesis   genetics   MeSH
• Tumor Microenvironment MeSH
• Neoplasm Proteins biosynthesis   genetics   MeSH
• Head and Neck Neoplasms genetics   pathology   MeSH
• Prognosis * MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Neoplasm Staging MeSH
• Gene Expression Profiling MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Biomarkers, Tumor MeSH
• Neoplasm Proteins MeSH

ABSTRACT: Quantum dots (QDs) are fluorescence nanoparticles (NPs) with unique optic properties which allow their use as probes in chemical, biological, immunological, and molecular imaging. QDs linked with target ligands such as peptides or small molecules can be used as tumor biomarkers. These particles are a promising tool for selective, fast, and sensitive tagging and imaging in medicine. In this study, an attempt was made to use QDs as a marker for human metallothionein (MT) isoforms 1 and 2. Four kinds of CdTe QDs of different sizes bioconjugated with MT were analyzed using the chip-CE technique. Based on the results, it can be concluded that MT is willing to interact with QDs, and the chip-CE technique enables the observation of their complexes. It was also observed that changes ranging roughly 6-7 kDa, a value corresponding to the MT monomer, depend on the hydrodynamic diameters of QDs; also, the MT sample without cadmium interacted stronger with QDs than MT saturated with cadmium. Results show that MT is willing to interact with smaller QDs (blue CdTe) rather than larger ones QDs (red CdTe). To our knowledge, chip-CE has not previously been applied in the study of CdTe QDs interaction with MT.

• Keywords
• Bioconjugation, Biomarkers, Chip-CE, Health effects, Metallothionein, Quantum dots,
• Publication type
• Journal Article MeSH