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Methods in Molecular Biology (Clifton,... 2021Mitochondria possess a genome that codes for proteins, in the same fashion as the nuclear genome. However, the small, circular mitochondrial DNA (mtDNA) molecule has a...
Mitochondria possess a genome that codes for proteins, in the same fashion as the nuclear genome. However, the small, circular mitochondrial DNA (mtDNA) molecule has a reduced base pair content, for it can only code for 2 rRNA, 22 tRNA molecules, and 13 proteins, all of them part of the mitochondrial respiratory chain. As such, all of the other mitochondrial components derive from nuclear genome. This separation leads to a requirement for a well-tuned coordination between both genomes, in order to produce fully functional mitochondria. A vast number of pathologies have been demonstrated to involve, to some extent, alterations in mitochondrial function that, no doubt, can be caused by alterations to the respiratory chain activity. As such, several methods and techniques have been developed to assess both content and function of mitochondrial proteins, in order to help understand mitochondrial involvement on the pathogenesis of disease. In this chapter, we will address some of these methods, with the main focus being on isolated mitochondria.
Topics: Animals; Cell Fractionation; Centrifugation; Electron Transport Chain Complex Proteins; Mice; Mitochondria, Liver; Mitochondrial Proteins; Oxidative Phosphorylation; Rats
PubMed: 34095995
DOI: 10.1007/978-1-0716-1433-4_2 -
Methods in Molecular Biology (Clifton,... 2021Platelet mitochondria can be used in the study of mitochondrial dysfunction in various complex diseases and can help in finding biological markers for diagnosing the...
Platelet mitochondria can be used in the study of mitochondrial dysfunction in various complex diseases and can help in finding biological markers for diagnosing the disease, monitoring its course and the effects of treatment. The aim of this chapter was to describe in detail the method of measuring mitochondrial respiration in platelets using high-resolution respirometry. The described method was successfully used for the study of mitochondrial dysfunction in neuropsychiatric diseases.
Topics: Blood Platelets; Cell Respiration; Humans; Mitochondria; Polarography
PubMed: 34080156
DOI: 10.1007/978-1-0716-1270-5_16 -
Journal of Analytical Methods in... 2021[This corrects the article DOI: 10.1155/2020/3646712.].
Erratum to "A Novel Method for Extraction of Galegine by Molecularly Imprinted Polymer (MIP) Technique Reinforced with Graphene Oxide and Its Evaluation Using Polarography".
[This corrects the article DOI: 10.1155/2020/3646712.].
PubMed: 33628574
DOI: 10.1155/2021/3548023 -
Journal of the Science of Food and... Sep 2021Lead (Pb ) is one of the most toxic heavy metals and can be found in various quantities in the environment. The five native probiotic bacteria and inulin were used to...
BACKGROUND
Lead (Pb ) is one of the most toxic heavy metals and can be found in various quantities in the environment. The five native probiotic bacteria and inulin were used to assess in vitro lead nitrate and lead acetate binding capacities, as well as removal potentials.
RESULTS
The highest decrease in media pH was seen for samples containing a combination of Lactobacillus paracasei IRBC-M 10784, lead nitrate and inulin (5.30 ± 0.012). The presence of inulin in the environment accelerated decreases in the pH of all samples with no significance. In all groups, lead nitrate-containing samples included maximum pH decreases. From the highest to the lowest, the ability of lead removal was linked to Lactobacillus acidophilus PTCC-1932 (88.48%), Bifidobacterium bifidum BIA-7 (85.32%), Bifidobacterium lactis BIA-6 (85.24%), Lactobacillus rhamnosus IBRC-M 10782 (83.18%) and L. paracasei IRBC-M 10784 (80.66%). Most species included the highest decrease in lead nitrate. Fourier-transform infrared spectroscopy (FTIR) analysis demonstrated that various functional groups (hydroxyl, carboxylic, carbonyl, amino and amide binds) on the bacterial cell wall were involved in lead ion binding during incubation. Principal component analysis of the FTIR results showed differences with respect to treated groups and control groups.
CONCLUSION
The results obtained in the present study reveal that the simultaneous use of native probiotics and inulin can be an effective and safe approach for removing various toxic substances, especially Pb. © 2021 Society of Chemical Industry.
Topics: Adsorption; Bifidobacterium; Biodegradation, Environmental; Cell Wall; Hydrogen-Ion Concentration; Inulin; Lactobacillus; Lead; Nitrates; Organometallic Compounds; Probiotics
PubMed: 33608880
DOI: 10.1002/jsfa.11160 -
Acta Neuropsychiatrica Apr 2021The present study was designed to test the hypothesis that there is a reduction in the activity of the enzyme cytochrome c oxidase (Cox) in Alzheimer's disease (AD). (Comparative Study)
Comparative Study Meta-Analysis
OBJECTIVE
The present study was designed to test the hypothesis that there is a reduction in the activity of the enzyme cytochrome c oxidase (Cox) in Alzheimer's disease (AD).
METHODS
Systematic review of literature and meta-analysis were used with data obtained from the PubMed, Scopus, MEDLINE, Lilacs, Eric and Cochrane. The keywords were Alzheimer's AND Cox AND mitochondria; Alzheimer's AND Cox AND mitochondria; Alzheimer's AND complex IV AND mitochondria. A total of 1372 articles were found, 23 of them fitting the inclusion criteria. The data were assembled in an Excel spreadsheet and analysed using the RevMan software. A random effects model was adopted to the estimative of the effect.
RESULTS
The data shows a significant decrease in the activity of the Cox AD patients and animal models.
CONCLUSION
Cox enzyme may be an important molecular component involved in the mechanisms underlying AD. Therefore, this enzyme may represent a possible new biomarker for the disease as a complementary diagnosis and a new treatment target for AD.
Topics: Alzheimer Disease; Animals; Colorimetry; Electron Transport Complex IV; Humans; Mice; Mitochondria; Models, Animal; Polarography; Rats; Software; Spectrophotometry
PubMed: 33256871
DOI: 10.1017/neu.2020.43 -
Drug Research Apr 2021Lead is a hazardous heavy metal, which causes many problems in the human body. Unfortunately, recent reports showed that smugglers and opium sellers add lead to drugs... (Observational Study)
Observational Study
BACKGROUND
Lead is a hazardous heavy metal, which causes many problems in the human body. Unfortunately, recent reports showed that smugglers and opium sellers add lead to drugs during the production procedure in order to increase its weight and cost.
PURPOSE
The aim of this study was development of a rapid and accurate method for measurement of blood lead levels (BLL) in the oral and inhaled opiate abuser people.
METHODS
BLL in samples obtained from the oral and inhaled opium addicted patients referring to Sina Hospital in Tabriz, Iran, during 2017 was compared with healthy control group (N=15). The wet digestion method was used to prepare whole blood and Mercury Droplet Electrode Polarography (MDEP) method was utilized for measurement of the lead content of digested samples.
RESULTS
Results showed that there were significant differences between the BLL of samples obtained from oral (17.12±74.61 μg/dL, p<0.0003) and inhaled (19.33±2.257 μg/dL, p<0.0001) opium addicted groups in comparison with healthy control group (4.669±0.3367 μg/dL).
CONCLUSION
Based on the results of this study it was observed that BLL in opium addicted people needs to be measured as soon as possible. Furthermore, screening of blood lead concentrations in opium-addicted people with a rapid and accurate MDEP method is very necessary and important.
Topics: Adolescent; Adult; Aged; Drug Contamination; Electrodes; Feasibility Studies; Female; Humans; Iran; Lead; Lead Poisoning; Male; Mercury; Middle Aged; Opium; Opium Dependence; Polarography; Sensitivity and Specificity; Young Adult
PubMed: 33184808
DOI: 10.1055/a-1293-6924 -
Applied Optics Sep 2020We examine the impact of illumination, aperture, and sample thickness on two division-of-focal-plane (DoFP) polarimeters, one created using a standard 3 T pixel and the...
We examine the impact of illumination, aperture, and sample thickness on two division-of-focal-plane (DoFP) polarimeters, one created using a standard 3 T pixel and the other with a forward-biased, logarithmic pixel. Across all measured metrics the logarithmic DoFP polarimeter was better able to track real-time changes in collagen alignment than the standard DoFP polarimeter.
Topics: Animals; Cattle; Collagen; Equipment Design; Polarography; Sensitivity and Specificity; Tendons
PubMed: 32976451
DOI: 10.1364/AO.398362 -
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... 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 -
Frontiers in Oncology 2020Lactic acidosis (3 to 40 mM, pH < 6.9) is a condition found in solid tumors because tumor cells have a high rate of glucose consumption and lactate production even in...
Lactic acidosis (3 to 40 mM, pH < 6.9) is a condition found in solid tumors because tumor cells have a high rate of glucose consumption and lactate production even in the presence of oxygen; nevertheless, the microenvironment might still provide a sufficient glucose supply. Lactic acidosis has been proposed to shift metabolism from aerobic glycolysis toward oxidative phosphorylation (OXPHOS). We tested if lung tumor cells cultured under lactic acidosis shift their metabolism from glycolysis to OXPHOS by consuming extracellular lactate, increasing growth rate. We analyzed lung adenocarcinoma (A-549, A-427) cell lines and non-transformed fibroblast cells (MRC-5), which were cultured using RPMI-1640 medium initially containing lactate (2 mM) and glucose (10 mM), at pH 7.2 or 6.2 and oxygen tension 21% O (normoxia) or 2% O (hypoxia). We obtained growth curves, as well as glucose consumption and lactate production rates (measured during exponential growth) for each cell line. HIF-1α (Hypoxia-inducible factor 1 α), CS (citrate synthase) and AMPK (AMP-activated protein kinase) transcript levels were analyzed using RT-qPCR. By flow cytometry, we determined: (a) expression of glucose transporters (GLUT)1 and 4; (b) lactate transporters (MCT)1 and 4; (c) cell cycle profile, and (d) protein levels of HIF-1α, total and phosphorylated AMPK (pAMPK). Mitochondrial functionality was evaluated by measuring O consumption in tumor cells using polarography and a Clark-type electrode. Tumor and non-transformed cells used both aerobic glycolysis and OXPHOS for obtaining energy. As of 48 h of culture, lactate levels ranged from (4.5-14 mM), thus forming a lactic environment. Lactic acidosis diminished GLUT1/GLUT4 expression and glucose consumption in A-549, but not in A-427 cells, and induced differential expression of HIF-1α, AMPK, and CS transcripts. A-427 cells increased pAMPK and HIF-1α levels and shifted their metabolism increasing OXPHOS; thus supporting cell growth. Conversely, A-549 cells increased HIF-1α protein levels, but did not activate AMPK and diminished OXPHOS. A-549 cells survived by arresting cells in G1-phase. Our findings show that lactic acidosis diminishes Warburg effect in tumor cells, but this change does not necessarily promote a shift to OXPHOS. Hence, lung adenocarcinomas show a differential metabolic response even when they are under the same microenvironmental conditions.
PubMed: 32596143
DOI: 10.3389/fonc.2020.00807