Extremely-low frequency magnetic field effects on sulfate reducing bacteria viability.

50 Hz magnetic fields effects on Sulfate Reducing Bacteria (SRB) viability were studied by colony forming units (CFU) counting. We found a 15% decrease of CFU number after magnetic field exposure (B=7.1 mT, f=50 Hz, t=24 min) compared to the control samples. These results are in good agreement with our previous work on other bacterial strains. The magnetic field effects on SRB are relatively large for small magnetic fields. The data correlations have been subjected to a simple physical chemical analysis, yielding surprisingly large estimates for the characteristic magnetic reaction susceptibility, even when the entire bacterium is assumed to be the direct target of interaction of the magnetic ac fields for the exposures in the time range from 3-24 min.

50 Hz magnetic field effect on the morphology of bacteria.

Atomic force microscopy was used to distinguish changes in morphology of bacteria induced by 50 Hz 10 mT magnetic field exposure. It is known that alternating magnetic field exposure causes decrease of viability of different bacterial strains. Previously we found that the viability of rod-like bacteria exposed to magnetic field decreased twice more in comparison with the spherical ones. Motivated by this fact we carried out this study with bacterial cells of both shapes. We used Escherichia coli (rod-like) and Paracoccus denitrificans (spherical) bacteria. As a result we have not observed any change in bacterial morphology neither of rod-like nor of spherical bacteria after 1 h, 50 Hz and 10 mT magnetic field exposure.

Effects of ELF-EMF on brain proteins in mice.

Effect of electromagnetic low frequency fields was studied on mice. We analyzed level of protein in brain of mouse. The levels of c-Jun and c-Fos in brains were measured using Western-blot techniques. Female and male laboratory mice were exposed for 4 days to magnetic field (Bm = 2 mT, f = 50 Hz). The exposure took place in cylindrical coil at laboratory temperature. After the experiment they were sacrificed and the level of protein c-Jun and c-Fos in different parts of brain were estimated. The expression of c-Fos was not affected by magnetic field on the other hand the expression of c-Jun decreased after magnetic field exposure. The results did not depend on sex of mice.

H3K9 acetylation and radial chromatin positioning.

Histone variants and their epigenetic modifications determine genome function, particularly transcription. However, whether regulation of gene expression can be influenced by nuclear organization or vice versa is not completely clear. Here, we analyzed the effect of epigenetic changes induced by a histone deacetylase inhibitor (HDACi) on the nuclear radial rearrangement of select genomic regions and chromosomes. The HDACi, sodium butyrate (NaBt), induced differentiation of human adenocarcinoma HT29 cells as well as a genome-wide increase in H3K9 acetylation. Three-dimensional analysis of nuclear radial distributions revealed that this increase in H3K9 acetylation was often associated with a repositioning of select loci and chromosomes toward the nuclear center. On the other hand, many centromeres resided sites more toward the nuclear periphery, similar to sites occupied by chromosome X. In more than two-thirds of events analyzed, central nuclear positioning correlated with a high level of H3K9 acetylation, while more peripheral positioning within interphase nuclei correlated with a lower level of acetylation. This was observed for the gene-rich chromosomes 17 and 19, TP53, and CCND1 genes as well as for gene-poor chromosome 18, APC gene, regions of low transcriptional activity (anti-RIDGEs), and the relatively transcriptionally less active chromosome X. These results are consistent with a role for epigenetic histone modifications in governing the nuclear radial positioning of genomic regions during differentiation.

Epigenome and chromatin structure in human embryonic stem cells undergoing differentiation.

Epigenetic histone (H3) modification patterns and the nuclear radial arrangement of select genetic elements were compared in human embryonic stem cells (hESCs) before and after differentiation. H3K9 acetylation, H3K9 trimethylation, and H3K79 monomethylation were reduced at the nuclear periphery of differentiated hESCs. Differentiation coincided with centromere redistribution, as evidenced by perinucleolar accumulation of the centromeric markers CENP-A and H3K9me3, central repositioning of centromeres 1, 5, 19, and rearrangement of other centromeres at the nuclear periphery. The radial positions of PML, RARalpha genes, and human chromosomes 10, 12, 15, 17, and 19 remained relatively stable as hESCs differentiated. However, the female inactive H3K27-trimethylated X chromosome occupied a more peripheral nuclear position in differentiated cells. Thus, pluripotent and differentiated hESCs have distinct nuclear patterns of heterochromatic structures (centromeres and inactive X chromosome) and epigenetic marks (H3K9me3, and H3K27me3), while relatively conserved gene density-related radial chromatin distributions are already largely established in undifferentiated hES cells.

Single-cell c-myc gene expression in relationship to nuclear domains.

Nuclear locations of the c-myc gene and its transcripts (c-myc (T)) have been investigated in relation to nuclear domains involved in RNA synthesis and processing. Transcription of the c-myc gene appears to be linked to the late G(1)- and preferentially to S-phases of the cell cycle. The c-myc gene and its transcripts were positioned non-randomly within the interphase nucleus; additionally, c-myc RNA signals accumulated at nucleoli. Using oligo-probes, designed to exon II and exon III of the c-myc gene, single c-myc (T) was preferentially observed in human carcinoma HT29 and A549 cells. Conversely, human embryonal teratocarcinoma NTERA cells were characterized by the presence of multiple c-myc RNA signals located in both the nucleoli and nucleoplasm. When accumulated at nucleoli, c-myc (T) occupied the periphery of this organelle, though not those associated with the cultivation surface. In HT29 cells, approximately 80% of c-myc (T) co-localized with the RNAP II positive regions, so-called transcription factories. However, in approximately 20% of the cells with c-myc transcripts, the c-myc (T) was released from the site of synthesis, and was not associated with either transcription factories or SC35 domains. In approximately 60% of nuclei with c-myc (T), these signals were located in close proximity to the SC35 regions, but promyelocytic leukaemia bodies were associated with c-myc (T) only in approximately 20% of the nuclei. Taken together, c-myc RNA signals were positioned in the most internal parts of the cell nuclei preferentially associated with the nucleoli. Specific nuclear and nucleolar positioning probably reflects the kinetics of c-myc RNA metabolism.

Effects of activated and nonactivated platelet-rich plasma on proliferation of human osteoblasts in vitro.

PURPOSE: The purpose of this study was to evaluate the effect of progressively increasing concentrations of activated and nonactivated platelet-rich plasma (PRP) on proliferation of human osteoblasts in vitro. MATERIALS AND METHODS: Human osteoblasts (hFOB 1.19) obtained from the American Type Culture Collection (ATCC, Manassas, VA) were used in the experiment. PRP was obtained from a 28-year-old healthy male volunteer by means of a Haemonetics gradient density cell separator (Haemonetics, Munich, Germany). Human thrombin was used to activate PRP. Three independent experiments were conducted. Samples containing 10% (0.38x increase in platelet count), 25% (0.95x increase in platelet count), 50% (1.95x increase in platelet count), and 75% (2.86x increase in platelet count) of activated PRP and nonactivated PRP were prepared including controls. After culture periods of 24, 48, and 72 hours osteoblast proliferation was evaluated by counting the number of cells using a Multisizer 3 Coulter Counter (Beckman Coulter, Inc, Fullerton, CA). RESULTS: After 24, 48, and 72 hours of incubation, the number of cells in the control group (without PRP) was higher than that of cells in samples containing activated or nonactivated PRP. Osteoblasts with 10% activated PRP (0.38x increase in platelet count) had the highest viability of all samples containing PRP. CONCLUSIONS: Activated PRP resulted in higher proliferation of osteoblasts compared with nonactivated PRP at concentrations of 10% (0.38x increase in platelet count) and 25% (0.95x increase in platelet count) in culture. This study failed to show significant increases in proliferation of human osteoblasts treated with activated or nonactivated PRP compared with controls in vitro.

Low-frequency magnetic field effect on cytoskeleton and chromatin.

The effect of magnetic fields on the living systems is studied in vivo or in vitro in very broad spectrum of organisms, cells and tissues. The mechanism of their acting is not known until now. We studied low-frequency magnetic field effect on cytoskeleton and on the structure of chromatin in human cells. We used cell line of small lung carcinoma (A549) and the effects of magnetic field on cytoskeleton and higher-order chromatin structure were analyzed 96 h of magnetic field exposure. Magnetic field generated by the cylindrical soil was homogenous and the cells were cultivated at 37 degrees C in humidified atmosphere containing 5% CO(2). Magnetic field induction was B(m)=2 mT and the net frequency f=50 Hz. In such affected and control cells the F-actin was estimated using FITC-conjugated Phalloidin and mitochondria were studied using MitoTracker (Molecular Probes). Images of cytoskeleton and genetic loci were acquired using confocal microscopy and analysis was performed by FISH 2.0 software. Slight morphological changes of F-actin filaments and mitochondria were observed in affected cells and nuclear condensation was found. These effects could be related to the process of cell death apoptosis probably induced by magnetic field. The studies aimed at centromeric heterochromatin (9cen) did not show statistically significant changes. Therefore, we suggest that magnetic field has no influence on higher order chromatin structure but certain changes could be observed on the level of cytoskeleton. However, these statements need a thorough verification. Our preliminary experiments will be extended and the effect of magnetic field on another structures of cytoskeleton and cell nuclei will be further studied.

Effect of electromagnetic fields on the denitrification activity of Paracoccus denitrificans.

Enzymatic activity (denitrification) of Paracoccus denitrificans was estimated electrochemically by reduction of duroquinone (DQ). Graphite electrodes covered with whole bacterial cells behind a dialysis membrane were used for measurement. P. denitrificans reduce nitrate and/or nitrite under anaerobic conditions to nitrogen gas. DQ acts as an electron mediator. After donation of the electrons to the respiratory system of the bacteria, produced DQ is reduced to durohydroquinone on the electrode surface electrocatalytically. P. denitrificans were exposed to low-frequency magnetic field (10 mT, 50 Hz) for 24 min. In comparison with the control samples, the reduction peak of I-E curves that represent denitrification activity of the cells decreased significantly after magnetic field exposure. The decrease of the peak current was about 20%. The CFU-colony forming units-method was used to estimate the number of surviving bacteria. After 24 min exposure of 10 mT magnetic field P. denitrificans culture on electrode indicates 21% bacterial death.

Effects of low-frequency magnetic fields on the viability of yeast Saccharomyces cerevisiae.

A 50 Hz magnetic field effect on the growth of yeasts Saccharomyces cerevisae was studied. The cylindrical coil induced magnetic fields with inductions up to 10 mT. Duration of exposure varied up to 24 min. Exposure took place at laboratory temperature (24-26 degrees C) and the air ventilator maintained the temperature at the place of the sample. We measured the growth curves of yeasts in broth and we calculated the number of CFU (colony forming units) on solid soil. We found that magnetic field decreases the number of yeasts, and slowed down their growth. The result is similar to the experiments with bacteria E. coli, S. aureus and L. adecarboxylata. It seems that the magnetic fields kill a part of yeasts and the bigger part of them survives and continues in their growth.

Comparison of the low-frequency magnetic field effects on bacteria Escherichia coli, Leclercia adecarboxylata and Staphylococcus aureus.

This work studies biological effects of low-frequency electromagnetic fields. We have exposed three different bacterial strains-Escherichia coli, Leclercia adecarboxylata and Staphylococcus aureus to the magnetic field (t<30 min, B(m)=10 mT, f=50 Hz) in order to compare their viability (number of colony-forming units (CFU)). We have measured the dependence of CFU on time of exposure and on the value of the magnetic field induction B(m). Viability decreases with longer exposure time and/or higher induction B(m) for all strains, but the quantity of the effect is strain-dependent. The highest decrease of the viability and the biggest magnetic field effect was observed with E. coli. The smallest magnetic field effect appears for S. aureus. From the measurement of the growth dynamics we have concluded that the decrease of the CFU starts immediately after the magnetic field was switched on.

Effects of low-frequency magnetic fields on bacteria Escherichia coli.

The effects of low-frequency magnetic fields (Bm=2.7-10 mT, f=50 Hz, time of exposure t=0-12 min, laboratory temperature) on the viability and oxidoreductive activity of gram-negative bacteria Escherichia coli were investigated. The growth of these bacteria was negatively affected by such fields. We compared two experimental systems--solenoid [Sb. Lek. 99 (1998) 455] and a cylindrical spool--to find differences between nonhomogeneous and "more homogeneous" magnetic fields. We observed analogous effects in both experimental conditions. The growth curve of the exposed bacteria was lower than the control one. The ability of bacteria to form colonies decreased with increasing magnetic field intensity and with increasing time of exposure. The oxidoreductive activity was measured using reduction of a tetrazolium salt. The decrease in oxidoreductive activity with increasing time of exposure was observed, but the effect was due to a lower amount of bacteria surviving the exposure to the magnetic fields. The decrease in oxidoreductive activity and ability to form colonies were compared with the assumption that the effect of magnetic field is probably bactericidal.