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Quantitative evaluation of the parameters of bacterial photoreactivation after exposure to ultraviolet light and ionizing radiation.

The purpose was to compare quantitatively the parameters of photoreactivation of an ultraviolet (UV) light hypersensitive strain of Escherichia coli Bs-1 irradiated with UV light and ionizing radiation. In addition, to evaluate the influence of the different physical and chemical factors on the parameters of the photoreactivation kinetics of the bacterial cells exposed to ionizing radiation. Survival curves and kinetics of the photoreactivation were measured in E. coli Bs-1 cells exposed to UV light (254 nm) and ionizing radiations (gamma-rays of 137Cs, gamma-rays of 60Co and 25 MeV pulsed X-rays). A mathematical model describing the process of photoreactivation in terms of a decreasing effective dose was applied to the experimental data obtained here and that published by others to evaluate quantitatively the probability of photoreactivation and the irreversible component of the radiation damage. Both the rate and extent of photoreactivation decreased in the following order of inactivating agents: WUV light, pulsed X-ray beam, gamma-ray of 60Co and gamma-ray of 137Cs. However, the irreversible component of radiation damage increased with the same order of radiations whereas the probability of photoreactivation per unit time was independent of the kind of radiation. After exposure to 6 MeV photons, the parameters of photoreactivation were changed in the presence of caffeine or after irradiation in the presence of the radioprotective agent dithiothreitol. The independence of the probability of photoreactivation on the quality of radiation indicates the cells have the same ability to photoreactivate damage produced by different kinds of radiations and is an additional argument indicating that during ionizing radiation a UV-like damage can be produced. The decrease in the extent and the rate of photoreactivation with radiation quality is explained by the formation of irreversible damage rather than by the impairment of the photorecovery process itself. Chemical and physical factors influencing the relative contribution of ionization and excitation on the ionizing radiation effect could modify both the extent of the photoreactivation and the probability of the recovery per unit time. It is concluded that the mathematical approach used here may be useful to reveal some new relationships between the parameters of photoreactivation.

DNA Damage↗

Mitogen-activated protein kinase, ERK1/2, is essential for the induction of vascular endothelial growth factor by ionizing radiation mediated by activator protein-1 in human glioblastoma cells.

Vascular Endothelial Growth Factor (VEGF)/Vascular Permeability Factor plays an important role in angiogenesis and cell proliferation of cancer cells. Glioblastoma cells are most malignant and show resistance to radiation therapy inducing VEGF to cause angiogenesis and brain edema. In the present study, the regulatory mechanism of the expression of VEGF by ionizing radiation was studied in three human glioblastoma cells. Induction of VEGF mRNA by ionizing radiation was dependent on dose and incubation time. Activator protein-1 (AP-1) was activated by 10 Gy of ionizing radiation in 1 h in T98G glioblastoma cells on an electrophoretic mobility shift assay. We constructed chimeric genes containing various regions of the VEGF promoter gene and the coding region for chloramphenicol acetyltransferase (CAT) and transiently transfected them to T98G cells. CAT assay with the VEGF promoter gene containing an AP-1 site demonstrated that the promoter activity of the VEGF gene was enhanced by ionizing radiation. Immunological analysis of the activity of mitogen-activated protein kinase, ERK1/2, showed that this activity is up-regulated by ionizing radiation. These results suggest that ERK1/2 pathway is involved in the up-regulation of VEGF expression ionizing radiation mediated by AP-1, which may lead to further neovascularization and proliferation of glioblastoma cells resistant to radiation therapy.

Blotting, Northern↗

Polarity effect of the thimble-type ionization chamber at a low dose rate.

It is known that the current collected from an ionization chamber exposed to a constant radiation intensity changes in magnitude when the polarity of the collecting potential is reversed. It is called the polarity effect of the ionization chamber. There are many possible causes that induce the polarity effect and one of them can be a field distortion due to a potential difference between the guard electrode and the collector. We studied how much the polarity effect depends on the design of the electrodes in the thimble-type ionization chamber. Two thimble-type ionization chambers, which had different electrode structures, were designed and fabricated at KAERI. We calculated the field distortions due to the potential difference between the guard electrode and the collector for the two ionization chambers. MAXWELL and Garfield were employed to calculate the electron drift lines inside the chamber. The polarity effects of the two ionization chambers were measured, and they were consistent with the field calculation. We could conclude that the polarity effect is mostly induced from the field distortion due to the potential difference between the guard electrode and the collector in our experiment and it depends significantly on the design of the electrodes.

Calibration↗

Extensive diversity of ionizing-radiation-resistant bacteria recovered from Sonoran Desert soil and description of nine new species of the genus Deinococcus obtained from a single soil sample.

The ionizing-radiation-resistant fractions of two soil bacterial communities were investigated by exposing an arid soil from the Sonoran Desert and a nonarid soil from a Louisiana forest to various doses of ionizing radiation using a (60)Co source. The numbers of surviving bacteria decreased as the dose of gamma radiation to which the soils were exposed increased. Bacterial isolates surviving doses of 30 kGy were recovered from the Sonoran Desert soil, while no isolates were recovered from the nonarid forest soil after exposure to doses greater than 13 kGy. The phylogenetic diversities of the surviving culturable bacteria were compared for the two soils using 16S rRNA gene sequence analysis. In addition to a bacterial population that was more resistant to higher doses of ionizing radiation, the diversity of the isolates was greater in the arid soil. The taxonomic diversity of the isolates recovered was found to decrease as the level of ionizing-radiation exposure increased. Bacterial isolates of the genera Deinococcus, Geodermatophilus, and Hymenobacter were still recovered from the arid soil after exposure to doses of 17 to 30 kGy. The recovery of large numbers of extremely ionizing-radiation-resistant bacteria from an arid soil and not from a nonarid soil provides further ecological support for the hypothesis that the ionizing-radiation resistance phenotype is a consequence of the evolution of other DNA repair systems that protect cells against commonly encountered environmental stressors, such as desiccation. The diverse group of bacterial strains isolated from the arid soil sample included 60 Deinococcus strains, the characterization of which revealed nine novel species of this genus.

DNA, Bacterial↗

Regulation of tumor necrosis factor gene expression by ionizing radiation in human myeloid leukemia cells and peripheral blood monocytes.

Previous studies have demonstrated that ionizing radiation induces the expression of certain cytokines, such as TNF alpha/cachectin. However, there is presently no available information regarding the molecular mechanisms responsible for the regulation of cytokine gene expression by ionizing radiation. In this report, we describe the regulation of the TNF gene by ionizing radiation in human myeloid leukemia cells. The increase in TNF transcripts by x rays was both time- and dose-dependent as determined by Northern blot analysis. Similar findings were obtained in human peripheral blood monocytes. Transcriptional run-on analyses have demonstrated that ionizing radiation stimulates the rate of TNF gene transcription. Furthermore, induction of TNF mRNA was increased in the absence of protein synthesis. In contrast, ionizing radiation had little effect on the half-life of TNF transcripts. These findings indicate that the increase in TNF mRNA observed after irradiation is regulated by transcriptional mechanisms and suggest that production of this cytokine by myeloid cells may play a role in the pathophysiologic effects of ionizing radiation.

Cell Line↗

Antioxidative effects of melatonin in protection against cellular damage caused by ionizing radiation.

Ionizing radiation is classified as a potent carcinogen, and its injury to living cells is, to a large extent, due to oxidative stress. The molecule most often reported to be damaged by ionizing radiation is DNA. Hydroxyl radicals (*OH), considered the most damaging of all free radicals generated in organisms, are often responsible for DNA damage caused by ionizing radiation. Melatonin, N-acetyl-5-methoxytryptamine, is a well-known antioxidant that protects DNA, lipids, and proteins from free-radical damage. The indoleamine manifests its antioxidative properties by stimulating the activities of antioxidant enzymes and scavenging free radicals directly or indirectly. Among known antioxidants, melatonin is a highly effective scavenger of *OH. Melatonin is distributed ubiquitously in organisms and, as far as is known, in all cellular compartments, and it quickly passes through all biological membranes. The protective effects of melatonin against oxidative stress caused by ionizing radiation have been documented in in vitro and in vivo studies in different species and in in vitro experiments that used human tissues, as well as when melatonin was given to humans and then tissues collected and subjected to ionizing radiation. The radioprotective effects of melatonin against cellular damage caused by oxidative stress and its low toxicity make this molecule a potential supplement in the treatment or co-treatment in situations where the effects of ionizing radiation are to be minimized.

Animals↗

Ionizing radiation activates expression of FOXO3a, Fas ligand, and Bim, and induces cell apoptosis.

Genotoxic stress such as ionizing radiation can induce DNA damage and promote cell-cycle arrest or apoptosis through either a p53-dependent or -independent pathway. Recently, members of the FOXO Forkhead transcription factor family have been implicated in playing a role in both DNA repair and apoptosis in mammalian cells that promoted us to examine the role of FOXO transcription factors in ionizing radiation-induced apoptosis. Here, we show that ionizing radiation can promote FOXO3a (FKHRL1) transcriptional activity and protein expression level, and induce nuclear translocation of FOXO3a in Saos2, a p53-null osteosarcoma cell line. Ionizing radiation stimulates expression of apoptosis-inducing proteins such as Fas ligand and the Bcl-2 interacting mediator of cell death (Bim) leading to cellular apoptosis. The observed upregulation of proapoptotic genes and apoptosis in cells without p53 in response to ionizing radiation suggests a novel p53-independent mechanism underlying ionizing radiation-induced apoptosis in cancer cells.

Apoptosis↗

Increased hprt mutant frequencies in Brazilian children accidentally exposed to ionizing radiation.

We have examined the effects of ionizing radiation on somatic mutations in vivo, using the hprt clonal assay. The study was performed on blood samples obtained from children exposed during a radiological accident that happened in 1987, in Goiânia, Brazil. The group of children exposed to ionizing radiation includes six males and four females ranging in age from 6 to 14 years at the time of exposure. The radiation doses ranged from 15 to 70 cGy. A Brazilian control group, not exposed to ionizing radiation, was also analyzed under similar conditions. the mean hprt mutant frequency for the exposed group was 4.6 times higher than the control group, although the cloning efficiency from the exposed group was significantly reduced. Linear regression analysis of the mutant frequency and ionizing radiation dose did not show a significant relationship between these two parameters. However, a reliable inverse relationship was demonstrated when the regression analysis was performed with nonselective cloning efficiency and ionizing radiation dose. It was demonstrated that nonselective cloning efficiency diminishes as ionizing radiation dose increases. To correct mutant frequencies for clonal events, the clonal relationship between the hprt mutant clones was examined by T-cell receptor analysis. The majority of the mutants analyzed represented individual clones, thus validating the observed mutant frequencies.

Adolescent↗

Electron impact and chemical ionization mass spectrometry of steroidal spirolactones.

Electron impact and chemical ionization mass spectra are reported for several steroidal spirolactones and their TMS ethers. The electron impact spectra were characterized generally by low abundance molecular ions and large numbers of fragment ions. Methane chemical ionization spectra exhibited high intensity [M+H]+ and/or [M+H-H2O]+ or [M+H-TMSOH]+ ions with few other fragment ions. Ammonia chemical ionization spectra had intense [M+H]+ and/or [M+NH4]+ ions with a few fragment ions generally formed by loss of H2O or TMSOH from these parent ions. Ammonia chemical ionization gave intense parent ions even for polyhydroxy compounds and their TMS ethers in contrast to methane chemical ionization. The results of this study suggest that a combination of electron impact with ammonia chemical ionization mass spectrometry would offer the best techniques for detection and identification of these compounds in biological fluids.

Ammonia↗

Desorption ionization mass spectrometry of synthetic porphyrins.

The desorption ionization mass spectra are reported for two classes of synthetic porphyrins--'tailed' derivatives of deuteroporphyrin IX and meso-substituted derivatized tetraphenylporphines. The mass spectra were obtained by three desorption ionization techniques--fast atom bombardment (FAB), desorption chemical ionization (DCI) and 'in-beam' or desorption electron ionization (DEI). The emphasis of this study is to determine the usefulness of the three desorption ionization methods for the molecular weight and side-chain structure determinations of synthetic porphyrins. The molecular parent ions and the major fragmentation pathways are discussed in detail. The DEI method yielded the fewest mass spectra showing molecular parent ions from the compounds studied, whereas FAB gave the most. The major fragmentation pathways were compound dependent and similar for all three ionization methods; generally, those cleavages alpha and beta to carbonyl groups predominated. The FAB spectra showed strong molecular adduct ions in several spectra that could be attributed to an in situ coordination of iron-containing porphyrins with the thio-containing liquid matrix molecules.

Chemical Phenomena↗

Ionized hypomagnesemia in patients undergoing orthotopic liver transplantation: a complication of citrate intoxication.

Using a new ion-selective electrode, plasma concentration of ionized magnesium was measured in nine adult patients undergoing orthotopic liver transplantation. Baseline plasma ionized magnesium (IMg2+) concentration (0.49 +/- 0.07 mmol/L) was slightly below normal values (0.55-0.66 mmol/L, 95% CI): Six patients had ionized hypomagnesemia and two of these had total hypomagnesemia. Ionized IMg2+ concentration progressively decreased during the dissection (0.45 +/- 0.07 mmol/L, p < 0.05) and anhepatic stage (0.38 +/- 0.07 mmol/L, p < 0.05) and returned toward baseline values by 2 hours after graft reperfusion. Plasma ionized calcium levels and acid-base status were maintained within normal limits during surgery. Serum citrate concentration increased during the dissection (0.58 +/- 0.60 mmol/L) and anhepatic stages (1.18 +/- 0.78 mmol/L), the result of transfusion of citrate-rich blood products in the absence of adequate hepatic function, and gradually returned toward baseline values after graft reperfusion. IMg2+ concentration inversely correlated with the plasma citrate concentration (r2 = 0.54). The results of this study demonstrate that ionized hypomagnesemia invariably occurs during liver transplantation and suggest that this derangement may be a clinical concern, because magnesium is an important cofactor for the maintenance of cardiovascular homeostasis. The data further suggest the clinical importance of supplementation with magnesium based on the monitoring of plasma IMg2+ concentration.

Analysis of Variance↗

New trends in the application of electron ionization to liquid chromatography-mass spectrometry interfacing.

I. Introduction 88 II. Cap-EI Interface 90 A. Interface Performance 92 III. Direct-EI Interface 93 A. Interface Performance 97 IV. Conclusions 103 Acknowledgments 103 References 103 --Two recent approaches for coupling capillary scale liquid chromatography and electron ionization mass spectrometry are reviewed and discussed. The first one, Cap-EI, is the latest evolution of the micro-scale particle beam interface, in which the nebulizer has been optimized to overcome the limitations of the former approach, in terms of sensitivity and linearity. It can be easily hosted in pre-existing instruments without major modifications and can use helium and the less-expensive nitrogen to generate library-matchable electron ionization spectra. The second one is a miniaturized interface for nano- and micro-HPLC, in which the interfacing process takes place into a suitably modified ion source. Because the eluate from the column is completely transferred into the ion source for ionization, superior sensitivity, linearity, and reproducibility are obtained. No signs of chemical ionization are observed at flow rates up to 1.5 microL/min. These two interfaces demonstrate that electron ionization can be successfully used for the analysis of small-medium molecules of various polarities, and also at the trace level. The possibility to record library-matchable electron ionization spectra offers the analyst a powerful tool that can be particularly useful in real-world applications.

Caffeine↗

Surface-activated no-discharge atmospheric pressure chemical ionization.

A new ionization method named surface-activated chemical ionization (SACI) has been realized. In this invention a commercially available atmospheric pressure chemical ionization (APCI) chamber, employed without any corona discharge (no-discharge APCI), has been modified with the insertion of a gold surface, leading to a significant improvement in the ionization efficiency. The ionization of the sample takes place by both gas-phase and surface-activated processes. This new ionization source is able to generate ions with high molecular mass and low charge states, leading to improved sensitivity and reduced noise. The new device has been tested in the analysis of some peptides. A comparison between the performance with and without the presence of the surface, and the optimization of the operating conditions (nebulizing gas flow, sample solution flow, pH of solution, and surface area), are reported and discussed.

Air Pressure↗

Anisole, a new dopant for atmospheric pressure photoionization mass spectrometry of low proton affinity, low ionization energy compounds.

Atmospheric pressure photoionization (APPI) is a novel method of ionization in liquid chromatography/mass spectrometry (LC/MS). It was originally developed in order to broaden the range of LC/MS ionizable compounds towards less polar compounds that cannot be analyzed by electrospray (ESI) and atmospheric pressure chemical ionization (APCI). Studies done thus far have shown that non-polar compounds that earlier were not ionizable in LC/MS can indeed be ionized by the use of APPI. However, the best ionization efficiency for low polarity samples has been achieved with low proton affinity (PA) solvents that are not suitable in reversed-phase LC (RP-LC). Here it is demonstrated that the signals for analytes with low proton affinities in acetonitrile can be increased 100-fold by using anisole as the dopant for APPI, which takes the sensitivity to the same level achieved in the analysis of high PA analytes.

Acetonitriles↗

Ionization energies of LinX (n = 2, 3; X = Cl, Br, I) molecules.

Molecules of Li(n)X (n = 2, 3; X = Cl, Br, I) were examined with a magnetic sector mass spectrometer by surface ionization using a triple rhenium filament impregnated with fullerene (C60). The ionization energies obtained for Li(2)Cl, Li(2)Br and Li(2)I molecules are 3.8 +/- 0.1, 3.9 +/- 0.1 and 4.0 +/- 0.1 eV, respectively. The first ionization energy of Li(2)Cl is documented, while there are no literature data for the ionization energies of Li(2)Br and Li(2)I. The molecules of Li(3)Cl, Li(3)Br and Li(3)I were detected experimentally for the first time with ionization energies of 4.0 +/- 0.1, 4.1 +/- 0.1 and 4.1 +/- 0.1 eV, respectively. The ionization energies of Li(n)X (n = 2, 3; X = Cl, Br, I) are in correlation with the theoretical prediction of their hyperlithiated configurations.

Journal Article↗

Plasma ionized calcium in brain-dead patients.

BACKGROUND: The mechanism of brain death-induced myocardial dysfunction remains debatable. Hypocalcemia is known to induce reversible myocardial dysfunction. However, the incidence of hypocalcemia and its effect on myocardial function during brain death is unknown. METHODS: In 54 consecutive brain-dead patients, we measured plasma total and ionized calcium concentrations, QT and corrected QT intervals, and left ventricular ejection fraction area (LVEFa), using transesophageal echocardiography. RESULTS: 49 (91%) of brain-dead patients had a decrease in total plasma total calcium concentration but only 19 (35%) had a decrease in plasma ionized calcium. Corrected total plasma calcium failed to predict ionized calcium concentration and QT intervals were not significantly different in normo and hypocalcemic patients. The LVEFa was not significantly different between normo and hypocalcemic patients (53 +/- 13 versus 50 +/- 20%), and no correlation was found between LVEFa and ionized calcium (R = 0.02, NS). Hypocalcemic patients required greater doses of dopamine (8.2 +/- 5.2 versus 5.0 +/- 3.4 micrograms.kg-.min-1, p < 0.02) to maintain arterial pressure. Hypocalcemia was associated with a higher volume loading and a lower plasma protide concentration which reflected hemodilution. CONCLUSION: A decrease in plasma ionized calcium is not frequent, rarely severe, and probably not the main mechanism of myocardial dysfunction in brain-dead patients. Hypocalcemic patients required higher doses of dopamine, suggesting a decrease in systemic resistance. Only direct measurement of ionized calcium can assess plasma calcium ion status in brain-dead patients.

Adult↗

Plasma ionized calcium during paediatric anaesthesia: effects of pH and succinylcholine.

Variations of plasma ionized calcium and pH were studied in 16 children anaesthetized with halothane and succinylcholine for minor surgery. During anaesthesia, a close negative correlation was noted between pH and ionized calcium. Intubation was followed by a fall of the ionized calcium concentration. A mean decrease of 0.04 mmol X L-1 was noted, without pH change. The acute elevation of carbon dioxide following extubation allowed the in vivo effects of pH on ionized calcium to be studied. For each pH unit decrease, a mean rise of 0.60 mmol X L-1 of ionized calcium was found. During the study, no significant variations of plasma total calcium concentration were observed. The variations in the hydrogen ion concentration and the administration of succinylcholine were accompanied by changes of the free calcium ion concentration, due probably to alterations in calcium binding to proteins, and perhaps to the formation of a calcium complex with anions. Our results emphasize the importance of knowledge of the pH for the interpretation of ionized calcium variations in the presence of acid-base disturbances.

Anesthesia, Inhalation↗

In vivo and in vitro ionized calcium variations induced by acute respiratory acid base disturbances.

The effects of acute respiratory acid base changes on the ionized calcium concentration were studied in mechanically ventilated patients and compared to the results which were found using an in vitro model. In both in vivo and in vitro studies, the ionized calcium changes correlated negatively with the pH variations. The good agreement between the in vivo and in vitro investigations showed a mean ionized calcium change of 0.40 mmol/l per pH unit. At the same time, the ionized calcium changes correlated positively with the changes of plasma bicarbonate. The mean ionized calcium change was 0.01 mmol/l per mmol/l bicarbonate variation. During respiratory acid base disturbances, the ionized calcium variations due to the competitive albumin binding between hydrogen and calcium ions are buffered and blunted by the bicarbonate ions.

Acid-Base Imbalance↗