The mechanism of toxicity and cellular response to metal ions present in the environment is still a very current area of research. In this work, which is a continuation of the study of the toxicity ...of metal ions released by fixed orthodontic appliances, eluates of archwires, brackets, ligatures, and bands are used to test the prooxidant effect, cytotoxicity, and genotoxicity on cell lines of the gastrointestinal tract. Eluates obtained after three immersion periods (3, 7, and 14 days) and with known amounts and types of metal ions were used. Four cell lines-CAL 27 (human tongue), Hep-G2 (liver), AGS (stomach) and CaCo-2 (colon)-were treated with each type of eluate at four concentrations (0.1×, 0.5×, 1.0×, and 2.0×) for 24 h. Most eluates had toxic effects on CAL 27 cells over the entire concentration range regardless of exposure time, while CaCo-2 proved to be the most resistant. In AGS and Hep-G2 cells, all samples tested induced free radical formation, with the highest concentration (2×) causing a decrease in free radicals formed compared to the lowest concentrations. Eluates containing Cr, Mn, and Al showed a slight pro-oxidant effect on DNA (on plasmid φX-174 RF I) and slight genotoxicity (comet assay), but these effects are not so great that the human body could not "resist" them. Statistical analysis of data on chemical composition, cytotoxicity, ROS, genotoxicity, and prooxidative DNA damage shows the influence of metal ions present in some eluates on the toxicity obtained. Fe and Ni are responsible for the production of ROS, while Mn and Cr have a great influence on hydroxyl radicals, which cause single-strand breaks in supercoiled plasmid DNA in addition to the production of ROS. On the other hand, Fe, Cr, Mn, and Al are responsible for the cytotoxic effect of the studied eluates. The obtained results confirm that this type of research is useful and brings us closer to more accurate in vivo conditions.
Despite numerous studies on ion release from orthodontic appliances, no clear conclusions can be drawn due to complex interrelations of multiple factors. Therefore, as the first part of a ...comprehensive investigation of cytotoxicity of eluted ions, the objective of this study was to analyze four parts of a fixed orthodontic appliance. Specifically, NiTi archwires and stainless steel (SS) brackets, bands, and ligatures were immersed in artificial saliva and studied for morphological and chemical changes after 3-, 7-, and 14-day immersion, using the SEM/EDX technique. Ion release profiles were analyzed for all eluted ions using inductively coupled plasma mass spectrometry (ICP-MS). The results demonstrated dissimilar surface morphologies among parts of the fixed appliance, due to variations in manufacturing processes. The onset of pitting corrosion was observed for the SS brackets and bands in the as-received state. Protective oxide layers were not observed on any of the parts, but adherent layers developed on SS brackets and ligatures during immersion. Salt precipitation, mainly KCl, was also observed. ICP-MS proved to be more sensitive than SEM/EDX and exhibited results undetected by SEM/EDX. Ion release was an order-of-magnitude higher for SS bands compared to other parts, which was attributed to manufacturing procedure (welding). Ion release did not correlate with surface roughness.
Endometriosis is a chronic inflammatory disease that leads to a series of pathological reactions. The basis is a changed proinflammatory activated immune system, which results in more pronounced ...oxidative stress, disturbed function of proteolysis and cell apoptosis. These processes are crucial in the development of the disease because their dysfunctional activities cause the progression of the disease. It is believed that the proteins excreted in the urine interact with each other and promote pathological processes in endometriosis.
We analyzed the urine proteome of patients and aimed to detect a potential protein biomarker for endometriosis in the urine proteome. We collected urine samples from 16 patients with endometriosis and 16 patients in the control group with functional ovarian cysts. The diagnosis for all patients was confirmed through pathohistological analysis. After the preanalytical preparation of the urine, chromatography and mass spectrometry (LC-MS/MS) used the technology of urine proteome analysis.
The main finding was a significantly different concentration of 14 proteins in the urine samples. We recorded a considerably higher concentration of proteins that have a significant role in activating the immune system (SELL), iron metabolism (HAMP) and cell apoptosis (CHGA) in endometriosis compared to controls. Proteins having an antioxidant function (SOD1) and a role in proteolysis of the extracellular matrix (MMP-9) were significantly reduced in endometriosis compared to controls.
Consistent with the known pathogenesis of endometriosis, the study results complement the pathological responses that occur with disease progression.
The treatment with fixed orthodontic appliances could have an important role in the induction of oxidative stress and associated negative consequences. Because of the simultaneous effects of ...corrosion, deformation, friction, and mechanical stress on fixed orthodontic appliances during treatment, degradation of orthodontic brackets and archwires occurs, causing higher concentrations of metal ions in the oral cavity. Corroded appliances cause the release of metal ions, which may lead to the increased values of reactive oxygen species (ROS) due to metal-catalyzed free radical reactions. Chromium, iron, nickel, cobalt, titanium, and molybdenum all belong to the group of transition metals that can be subjected to redox reactions to form ROS. The estimation of health risk due to the amount of heavy metals released and the level of selected parameters of oxidative stress generated for the time of treatment with fixed orthodontic appliances is presented. Approaches to avoid oxidative stress and recommendations for the preventive use of topical or systemic antioxidants during orthodontic treatment are discussed.
The yeast Saccharomyces cerevisiae is a useful model for studying the influence of different stress factors on eukaryotic cells. In this work we used the pesticide imidacloprid, in the Confidor ...formulation, as the stress factor and analyzed its influence on the metabolic activity, proteome and lipid content and composition of Saccharomyces cerevisiae yeast. During the cultivation of yeast, the lowest recommended application dose of Confidor (0.025%, v/v) was added to the growth media and its influence on the mitochondria, cytosol with microsomes, and the whole yeast cells was monitored. The results show that under the stress provoked by the toxic effects of Confidor, yeast cells density significantly decreased and the percentage of metabolically disturbed cells significantly increased comparing with untreated control. Also, there was a downregulation of majority of glycolytic, gluconeogenesis, and TCA cycle enzymes (Fba1, Adh1, Hxk2, Tal1, Tdh1,Tdh3, Eno1) thus providing enough acetyl-CoA for the lipid restructuring and accumulation mechanism since we have found the changes in the cell and mitochondrial lipid content and FA composition. This data suggest that lipids could be the molecules that orchestrate the answer of the cells in the stress response to the Confidor treatment.
In the pathogenesis of endometriosis, a number of pathological reactions occur. Proteins secreted in the urine are thought to interact with each other and stimulate the pathological processes in ...endometriosis. Identifying one or more proteins that are specific enough and could serve as biomarkers for endometriosis is both a challenge and a necessity that would facilitate diagnosis. The urine of patients treated in a tertiary university hospital between July 1, 2020 and June 30, 2021 was analyzed. The studied group consists of patients who were treated surgically for endometriosis and in whom the diagnosis was confirmed by pathohistological analysis. The control group consists of patients who were operated for functional ovarian cysts. Urinary proteins were analyzed by chromatography and mass spectrometry (LC-MS/MS). We identified 17 proteins in urine whose concentrations were statistically significantly different in the group with endometriosis (N = 16) compared with the control groups (N = 16). The detected proteins were classified into groups according to their function in invasion, migration and proliferation, proteolysis, immune system, cell adhesion and vascular system. For all mentioned proteins the difference in concentration is statistically significant p < 0.005. Proteins are secreted in the urine of patients with endometriosis that may be involved in the pathogenesis of the disease and are possible biomarkers for endometriosis.
•Proteins in urine; possible non-invasive biomarkers of endometriosis.•Oxidative stress in the peritoneal cavity promotes pathological reactions of endometriosis.•The chronic inflammatory state of endometriosis increases the expression of proteins in the urine.•Proteins with apoptosis function in endometriosis; increased secretion with less effect.
Yeast
Saccharomyces cerevisiae
is an ideal model organism for studying molecular mechanisms of the stress response provoked by metals. In this work, yeast cells response to iron (Fe
3+
) or lead (Pb
...2+
) exposure was tested and compared. Survival test was used to determine testing doses of metal ions—for Fe
3+
it was 4 mM and for Pb
2+
8 mM. These (high, over-loaded) doses provoked comparable values of growth inhibition, but different values in vitality measurement. The percentage of metabolically active cells, determined by fluorescent FUN-1 dye, was lower in Pb
2+
than in Fe
3+
treated cells. Besides, endogenous antioxidant defence systems in the cells treated with Pb
2+
were less efficient compared to Fe
3+
. At the mitochondrial level, the effects of metal ions were in correlation with the results of cell metabolic activity. The mitochondrial proteome of Pb
2+
treated cells showed the domination of protein downregulation. Yeast cells treated either with Fe
3+
or Pb
2+
shared 19 common significantly changed proteins. The affected proteins were involved in different cellular process and amongst them only five proteins belong to energy and carbohydrate metabolism, and protein biosynthesis. Based on all obtained results, it is possible to conclude that the effects of Fe
3+
and Pb
2+
on yeast cells show rather specific patterns of toxicity and stress response.
The antioxidant activity of three types of pumpkin seed oil or oil mixtures (cold- -pressed, produced from roasted seed paste and salad) produced in the northern part of Croatia and the kinetics of ...their behaviour as free radical scavengers were investigated using DPPH˙. In addition, the involvement of oil tocopherol isomers (α-, γ- and δ-) in different steps of DPPH˙ disappearance and their impact on the rate of reaction were analysed. The kinetics of DPPH˙ disappearance is a two-step process. In the first step, rapid disappearance of DPPH˙ occurs during the first 11 min of the reaction, depending on the oil type, followed by a slower decline in the second step. To describe DPPH˙ disappearance kinetics, six mathematical models (mono- and biphasic) were tested. Our findings showed that γ- and δ-tocopherols affected DPPH˙ disappearance during the first step, and α-tocopherol in the second step of the reaction. Moreover, α-tocopherol demonstrated 30 times higher antioxidant activity than γ- and δ-tocopherols. The results indicated the biphasic double-exponential behaviour of DPPH˙ disappearance in oil samples, due to the complexity of reactions that involve different tocopherol isomers and proceed through different chemical pathways.
The neutral lipid fraction of the aerobically grown starter yeast culture of a Saccharomyces cerevisiae brewing strain, and three-first recycled yeast generations exposed to multiple stress factors ...during beer fermentation was studied. No pronounced changes in the cellular neutral lipid content between the non-stressed starter and stressed recycled cells were found. However, it was found that recycled yeast generations modulate their neutral lipid composition during fermentation. The ergosterol content was increased at the expense of steryl esters (SEs) and squalene, which resulted in a higher ergosterol/SEs molar ratio and a slightly higher ergosterol/squalene molar ratio. In addition, the proportion of unsaturated fatty acids, mainly palmitoleic acid increased in the neutral lipid fraction of the stressed recycled yeast generations. These results suggest that some specific neutral lipid species and fatty acids stored in the neutral lipid fraction are involved in the adaptive response of the brewer's yeast to stressful fermentation conditions. The striking finding was a high squalene content in the neutral lipid fraction of both the starter yeast culture and recycled yeast generations (22.4 vs. 19-20%, respectively), implying a possible biotechnological exploitation of this biologically active molecule from the yeast biomass.
The disease in which we observe the invasion and growth of endometrial cells on extrauterine tissues and organs with the creation of a chronic inflammatory state is called endometriosis. It causes ...infertility and is present in more than 30% of patients with endometriosis. Diagnosis and treatment of the disease is most often delayed for about 8 years after the first symptoms were reported. The symptomatology of endometriosis is varied, and there is no non-invasive way of diagnosis, and this is the reason for the delayed start of treatment. The development of endometriosis activates pathological processes such as the invasion and proliferation of endometriotic cells, the formation of adhesions and the activation of the immune system, which result in increased protein expression. The aim of this research is to compare the concentrations of total proteins in the urine of subjects with endometriosis with those of the control group and possibly identify a biomarker for the diagnosis of endometriosis.
Prospective urine analysis of 141 patients who were hospitalized and surgically treated at the Clinic for Gynecology and Obstetrics of KBC Rijeka from 08/21/2021 until 07/30/2022. The urine of subjects with endometriosis (N=84) and without endometriosis (n=57) was analyzed.
Total protein in the urine is increased in the urine of subjects with endometriosis, but the total amount of protein does not correlate with the degree of disease progression.
An increase in the level of total proteins in urine in subjects with endometriosis is a possible non-invasive diagnostic biomarker. Patients with endometriosis are grouped after the concentration of total proteins greater than 5000 μg/μl.
PDF
