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International Journal of Molecular... Jul 2020For deep space exploration, reproductive health must be maintained to preserve the species. However, the mechanisms underlying the effect of changes in gravity on male...
For deep space exploration, reproductive health must be maintained to preserve the species. However, the mechanisms underlying the effect of changes in gravity on male germ cells remain poorly understood. The aim of this study was to determine the effect of simulated micro- and hypergravity on mouse sperm motility and the mechanisms of this change. For 1, 3 and 6 h, mouse sperm samples isolated from the caudal epididymis were subjected to simulated microgravity using a random position machine and 2g hypergravity using a centrifuge. The experimental samples were compared with static and dynamic controls. The sperm motility and the percentage of motile sperm were determined using microscopy and video analysis, cell respiration was determined by polarography, the protein content was assessed by Western blotting and the mRNA levels were determined using qRT-PCR. The results indicated that hypergravity conditions led to more significant changes than simulated microgravity conditions: after 1 h, the speed of sperm movement decreased, and after 3 h, the number of motile cells began to decrease. Under the microgravity model, the speed of movement did not change, but the motile spermatozoa decreased after 6 h of exposure. These changes are likely associated with a change in the structure of the microtubule cytoskeleton, and changes in the energy supply are an adaptive reaction to changes in sperm motility.
Topics: Animals; Cell Respiration; Cells, Cultured; Hypergravity; Male; Mice; Proteins; RNA, Messenger; Sperm Motility; Spermatozoa; Weightlessness; Weightlessness Simulation
PubMed: 32709012
DOI: 10.3390/ijms21145054 -
Antioxidants (Basel, Switzerland) Dec 2022Redox disbalance in placental cells leads to the hyperproduction of reactive oxygen species (ROS), it mediates the dysregulation of the maternal immune tolerance to a...
Interrelation between miRNAs Expression Associated with Redox State Fluctuations, Immune and Inflammatory Response Activation, and Neonatal Outcomes in Complicated Pregnancy, Accompanied by Placental Insufficiency.
Redox disbalance in placental cells leads to the hyperproduction of reactive oxygen species (ROS), it mediates the dysregulation of the maternal immune tolerance to a semi-allogenic fetus, inducing pro-inflammatory reactions, and it plays a central role in perinatal complications and neonatal disease programming. Microvesicles, which provide transplacental communication between a mother and fetus, contain microRNAs (miRNAs) that are sensitive to oxidative stress (OS) mediators and can control the balance of ROS production and utilization in target cells. In the context of this paradigm, we evaluated the markers of redox balance—MDA and 4-HNE for OS and GPx, and SOD, CAT, and GSH for the antioxidant system in the cord blood plasma of newborns diagnosed with fetal growth restriction (FGR)—by using polarography, spectrophotometry, and Western blotting. The expression of miRNAs associated with OS, immune and inflammatory responses in the blood plasma of newborns with intrauterine pneumonia (IP), neonatal sepsis (NS) and respiratory distress syndrome (RDS) was evaluated by a quantitative RT-PCR. Significant differences in the MDA level and reduced GPx and CAT activity were co-found for early-onset FGR (i.e., <34 gestational age). Significant correlations were found with a low birth weight by Apgar scores with reduced levels of antioxidant enzymes. Indeed, the level of OS markers increased in early-onset FGR in newborns with an extremely low body weight and high echogenicity of the periventricular zones, and reduced in late-onset FGR in newborns with IP, hyperbilirubinemia, intraventricular hemorrhage (IVH) and cerebral cysts. A prognostic model (AUC = 1; cutoff—0.5) was developed to assess the risk of IVH in newborns diagnosed with FGR based on the assessment of the OS markers (i.e., MDA + 4 HNE + CAT + GSH). A significant increase in the miR-127-3p expression was found in the plasma of newborns with NS (<32 GA; p ≤ 0.03 and >32 GA; p ≤ 0.009), IP (>32 GA; p ≤ 0.0001), and RDS (>32 GA; p ≤ 0.03). At the same time, the expression of miR-25-3p (p ≤ 0.03) was increased only in newborns with NS (>32 GA; p ≤ 0.03). The risk of developing IVH for premature newborns with IP (AUC = 0.8; cutoff—0.6) and NS (AUC = 0.68; cutoff—0.49) was assessed based on the miR-25-3p and miR-127-3p expression. Several key transcription factors were identified as the targets of studied miRNA since they are involved in the regulation of OS (NRF2), signaling and activation of the immune response (PRDM1, CCL26) and, also, inflammatory responses (NFKB1). The study of these miRNAs showed that they are involved in the modulation of processes leading to perinatal complications. Moreover, miR-127-3p is related to pro-inflammatory reactions and the formation of the macrophage phenotype in newborns with IP, NS, and RDS, while miR-25-3p is associated with an inhibition of macrophage migration and activation of antioxidant enzymes, which may prevent the development of oxidative damage in newborns with NS.
PubMed: 36670868
DOI: 10.3390/antiox12010006 -
American Journal of Physiology.... Aug 2019High energy expenditure is reported in cystic fibrosis (CF) animal models and patients. Alterations in skeletal muscle oxidative capacity, fuel utilization, and the...
High energy expenditure is reported in cystic fibrosis (CF) animal models and patients. Alterations in skeletal muscle oxidative capacity, fuel utilization, and the creatine kinase-phosphocreatine system suggest mitochondrial dysfunction. Studies were performed on congenic C57BL/6J and F508del () mice. Indirect calorimetry was used to measure gas exchange to evaluate aerobic capacity during treadmill exercise. The bioenergetic function of skeletal muscle subsarcolemmal (SSM) and interfibrillar mitochondria (IFM) was evaluated using an integrated approach combining measurement of the rate of oxidative phosphorylation by polarography and of electron transport chain activities by spectrophotometry. CF mice have reduced maximal aerobic capacity. In SSM of these mice, oxidative phosphorylation was impaired in the presence of complex I, II, III, and IV substrates except when glutamate was used as substrate. This impairment appeared to be caused by a defect in complex V activity, whereas the oxidative system of the electron transport chain was unchanged. In IFM, oxidative phosphorylation and electron transport chain activities were preserved, whereas complex V activity was reduced, in CF. Furthermore, creatine kinase activity was reduced in both SSM and IFM of CF skeletal muscle. The decreased complex V activity in SSM resulted in reduced oxidative phosphorylation, which could explain the reduced skeletal muscle response to exercise in CF mice. The decrease in mitochondrial creatine kinase activity also contributed to this poor exercise response.
Topics: Animals; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Energy Metabolism; Female; Male; Mice; Mice, Inbred C57BL; Mice, Inbred CFTR; Mice, Transgenic; Mitochondria, Muscle; Muscle, Skeletal; Oxidative Phosphorylation; Oxidative Stress; Physical Conditioning, Animal; Sequence Deletion
PubMed: 31211618
DOI: 10.1152/ajpendo.00064.2019 -
Journal of Trace Elements in Medicine... Jul 2024This study examines the imprecision of zinc (Zn) measurements across various clinical detection methods by analyzing the external quality assessment (EQA) data from 2018...
BACKGROUND
This study examines the imprecision of zinc (Zn) measurements across various clinical detection methods by analyzing the external quality assessment (EQA) data from 2018 to 2022. The findings of this study aim to offer recommendations for enhancing Zn measurements.
METHODS
Participating laboratories were grouped into peer categories based on the detection methods. The robust mean and coefficient of variation (CV) of the samples were calculated following ISO 13528 guidelines. The evaluation criteria for optimal, desirable, and minimum allowable imprecision in Zn estimation are 2.50%, 5.05%, and 7.55%, respectively, based on biological variation. Furthermore, the study examined inter-lab CVs, inter-method bias, and the passing rate. The impact of sample concentration on CVs and the pass rate was also investigated.
RESULTS
Over the past five years, 4283 laboratories participated in the EQA program, showing a high pass rate that improved as sample concentration increased. Differential pulse polarography (DPP) demonstrated stable and low CVs (0.61-1.86%). Although differential pulse stripping (DPS) was less stable than DPP, it still exhibited a low CV (0.71-3.10%). Graphite furnace atomic absorption spectrometry (GFAAS) and flame atomic absorption spectrometry (FAAS) performed similarly and displayed stable CVs (2.39-4.42%) within the acceptable range of desirable imprecision (5.05%). However, the CVs for ICP-MS were unacceptable in three out of the five years (5.28-6.20%). In 2022, the number of participating laboratories for DDP, DPS, GFAAS, FAAS and ICP-MS is 131, 35, 35, 820 and 72, respectively.
CONCLUSION
This study provides reliable insights into the imprecision of Zn measurements in clinical laboratories. The findings indicate that additional efforts are required to reduce the imprecision of ICP-MS in Zn measurements.
Topics: Zinc; China; Humans; Quality Control
PubMed: 38636293
DOI: 10.1016/j.jtemb.2024.127451 -
Ecotoxicology and Environmental Safety May 2023Heavy metals are ubiquitous environmental pollutants that are extremely dangerous for public health, but the molecular mechanisms of their cytotoxic action are still not...
Modulators of mitochondrial ATP-sensitive potassium channel affect cytotoxicity of heavy metals: Action on isolated rat liver mitochondria and AS-30D ascites hepatoma cells.
Heavy metals are ubiquitous environmental pollutants that are extremely dangerous for public health, but the molecular mechanisms of their cytotoxic action are still not fully understood. In the present work, the possible contribution of the mitochondrial ATP-sensitive potassium channel (mK(ATP)), which is usually considered protective for the cell, to hepatotoxicity caused by heavy metals was investigated using polarography and swelling techniques as well as flow cytometry. Using isolated liver mitochondria from adult male Wistar rats and various potassium media containing or not containing penetrating anions (KNO, KSCN, KAcet, KCl), we studied the effect of mK(ATP) modulators, namely its blockers (5-hydroxydecanoate, glibenclamide, ATP, ADP) and activators (diazoxide, malonate), on respiration and/or membrane permeability in the presence of hepatotoxins such as Cd, Hg, and Cu. It has been shown for the first time that, contrary to Hg and depending on media used, the mK(ATP) modulators affect Cd- and/or Cu-induced alterations in mitochondrial swelling and respiration rates, although differently, nevertheless, in the ways compatible with mK(ATP) participation in both these cases. On rat AS-30D ascites hepatoma cells, it was found that, unlike Cd, an increase in the production of reactive oxygen species was observed with the simultaneous use of Cu and diazoxide; in addition, there was no protective effect of diazoxide against cell death, which also occurred in the presence of Cu. In conclusion, the relationships (functional, structural and/or regulatory) between mK(ATP), components of the mitochondrial electron transport chain (CI, CII-CIII and/or ATP synthase, CV) and mitochondrial permeability transition pores were discussed, as well as the role of these molecular structures in the mechanisms of the cytotoxic action of heavy metals.
Topics: Rats; Male; Animals; Mitochondria, Liver; KATP Channels; Diazoxide; Cadmium; Ascites; Carcinoma, Hepatocellular; Rats, Wistar; Metals, Heavy; Mercury; Liver Neoplasms; Adenosine Triphosphate
PubMed: 36989557
DOI: 10.1016/j.ecoenv.2023.114829 -
Frontiers in Nutrition 2021Fatty acid overload, either of the saturated palmitic acid (PA) or the unsaturated oleic acid (OA), causes triglyceride accumulation into specialized organelles termed...
Fatty acid overload, either of the saturated palmitic acid (PA) or the unsaturated oleic acid (OA), causes triglyceride accumulation into specialized organelles termed lipid droplets (LD). However, only PA overload leads to liver damage mediated by mitochondrial dysfunction. Whether these divergent outcomes stem from differential effects of PA and OA on LD and mitochondria joint dynamics remains to be uncovered. Here, we contrast how both fatty acids impact the morphology and interaction between both organelles and mitochondrial bioenergetics in HepG2 cells. Using confocal microscopy, we showed that short-term (2-24 h) OA overload promotes more and bigger LD accumulation than PA. Oxygen polarography indicated that both treatments stimulated mitochondrial respiration; however, OA favored an overall build-up of the mitochondrial potential, and PA evoked mitochondrial fragmentation, concomitant with an ATP-oriented metabolism. Even though PA-induced a lesser increase in LD-mitochondria proximity than OA, those LD associated with highly active mitochondria suggest that they interact mainly to fuel fatty acid oxidation and ATP synthesis (that is, metabolically "active" LD). On the contrary, OA overload seemingly stimulated LD-mitochondria interaction mainly for LD growth (thus metabolically "passive" LDs). In sum, these differences point out that OA readily accumulates in LD, likely reducing their toxicity, while PA preferably stimulates mitochondrial oxidative metabolism, which may contribute to liver damage progression.
PubMed: 34869541
DOI: 10.3389/fnut.2021.775382 -
Molecules (Basel, Switzerland) Mar 2023The results of kinetic measurements revealed an accelerating effect of acetazolamide (ACT) on the multistep In(III) ions electroreduction in chlorates(VII) on a novel,...
Kinetics and Mechanism of In(III) Ions Electroreduction on Cyclically Renewable Liquid Silver Amalgam Film Electrode: Significance of the Active Complexes of In(III)-Acetazolamide.
The results of kinetic measurements revealed an accelerating effect of acetazolamide (ACT) on the multistep In(III) ions electroreduction in chlorates(VII) on a novel, cyclically renewable liquid silver amalgam film electrode (R-AgLAFE). The kinetic and thermodynamic parameters were determined by applying the DC polarography, square-wave (SWV) and cyclic voltammetry (CV), as well as electrochemical impedance spectroscopy (EIS). It was shown that ACT catalyzed the electrode reaction ("cap-pair" effect) by adsorbing on the surface of the R-AgLAFE electrode. The catalytic activity of ACT was explained as related to its ability to form active In(III)- acetazolamide complexes on the electrode surface, facilitating the electron transfer process. The active complexes constitute a substrate in the electroreduction process and their different structures and properties are responsible for differences in the catalytic activity. The determined values of the activation energy ΔH≠ point to the catalytic activity of ACT in the In(III) ions electroreduction process in chlorates(VII). Analysis of the standard entropy values ΔS0 confirm changes in the dynamics of the electrode process.
PubMed: 37049705
DOI: 10.3390/molecules28072942 -
Scientific Reports Sep 2023The present study aims to determine the relationship between the concentration of arsenic in the groundwater of Hamadan province and the mortality rate due to various...
The present study aims to determine the relationship between the concentration of arsenic in the groundwater of Hamadan province and the mortality rate due to various types of malignancies, congenital anomalies, diabetes mellitus and Alzheimer's. Mortality data due to various causes of death in Hamadan province were collected for five years (2016-2020). Sampling of drinking water was determined in the reference laboratory using polarography method. Poisson regression was used to investigate the relationship between arsenic level and the death rate due to various types of disease, at a significant level (p value < 0.05). According to the results of Poisson regression, among the various causes of death (N = 8042), Alzheimer's 5.94 (3.67-9.61), diabetes mellitus 4.05 (3.5-5.37), congenital malformations 2.98 (1.88-4.72), breast cancer 2.72 (1.56-4.71), leukemia 1.90 (1.24-2.92), stomach cancer 1.64 (1.28-2.10), Liver cancer 1.58 (1.58-2.30), other digestive organs 5.86 (3.38-10.16), meninges and brain cancer 1.57 (1.02-2.41) showed the highest relationship with arsenic contamination. The results of this study could be evidence for a positive and significant relationship between arsenic concentrations and mortality rates due to cancers, diabetes mellitus, Alzheimer disease, and congenital malformations. Therefore, it's necessary to use appropriate water treatment methods to remove arsenic at the source in contaminated areas.
Topics: Humans; Arsenic; Alzheimer Disease; Brain Neoplasms; Diabetes Mellitus; Leukemia
PubMed: 37726351
DOI: 10.1038/s41598-023-42744-4 -
International Journal of Molecular... Aug 2020The role of the Earth's gravitational and magnetic fields in the evolution and maintenance of normal processes of various animal species remains unclear. The aim of this...
The role of the Earth's gravitational and magnetic fields in the evolution and maintenance of normal processes of various animal species remains unclear. The aim of this work was to determine the effect of simulated microgravity and hypomagnetic conditions for 1, 3, and 6 h on the sperm motility of the fruit fly . In addition to the usual diet, the groups were administered oral essential phospholipids at a dosage of 500 mg/kg in medium. The speed of the sperm tails was determined by video recording and analysis of the obtained video files, protein content by western blotting, and cell respiration by polarography. The results indicated an increase in the speed of movement of the sperm tails after 6 h in simulated microgravity. The levels of proteins that form the axoneme of the sperm tail did not change, but cellular respiration was altered. A similar effect occurred with the administration of essential phospholipids. These results may be due to a change in the level of phosphorylation of motor proteins. Exposure to hypomagnetic conditions led to a decrease in motility after 6 h against a background of a decrease in the rate of cellular respiration due to complex I of the respiratory chain. This effect was not observed in the flies that received essential phospholipids. However, after 1 h under hypomagnetic conditions, the rate of cellular respiration also increased due to complex I, including that in the sperm of flies receiving essential phospholipids.
Topics: Administration, Oral; Animals; Cell Respiration; Drosophila Proteins; Drosophila melanogaster; Magnetic Fields; Male; Phospholipids; Sperm Motility; Spermatozoa; Weightlessness; Weightlessness Simulation
PubMed: 32825268
DOI: 10.3390/ijms21175985 -
International Journal of Molecular... Aug 2021Olivacine and ellipticine are model anticancer drugs acting as topoisomerase II inhibitors. Here, we present investigations performed on four olivacine derivatives in...
Olivacine and ellipticine are model anticancer drugs acting as topoisomerase II inhibitors. Here, we present investigations performed on four olivacine derivatives in light of their antitumor activity. The aim of this study was to identify the best antitumor compound among the four tested olivacine derivatives. The study was performed using CCRF/CEM and MCF-7 cell lines. Comet assay, polarography, inhibition of topoisomerase II activity, histone acetylation, and molecular docking studies were performed. Each tested compound displayed interaction with DNA and topoisomerase II, but did not cause histone acetylation. Compound (9-methoxy-5,6-dimethyl-1-({[1-hydroxy-2-(hydroxymethyl)butan-2-yl]amino}methyl)-6-pyrido[4,3-]carbazole) was found to be the best candidate as an anticancer drug because it had the highest affinity for topoisomerase II and caused the least genotoxic damage in cells.
Topics: Antineoplastic Agents; Cell Line, Tumor; Cell Survival; DNA; DNA Topoisomerases, Type II; Ellipticines; Humans; Molecular Docking Simulation; Neoplasms; Topoisomerase II Inhibitors; Tumor Cells, Cultured
PubMed: 34445198
DOI: 10.3390/ijms22168492